Macrocycles for use in treating disease

ABSTRACT

The present disclosure relates to certain chiral diaryl macrocyclic derivatives, pharmaceutical compositions containing them, and methods of using them to treat cancer.

CROSS REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.17/110,051, filed Dec. 2, 2020, which claims priority to U.S.Provisional Application No. 62/943,098, filed Dec. 3, 2019; and U.S.Provisional Application No. 63/015,937, filed Apr. 27, 2020, each ofwhich is incorporated herein in its entirety for all purposes.

TECHNICAL FIELD

The present disclosure relates to certain diaryl macrocyclicderivatives, pharmaceutical compositions containing them, and methods ofusing them to treat cancer.

BACKGROUND

Protein kinases are key regulators for cell growth, proliferation andsurvival. Genetic and epigenetic alterations accumulate in cancer cellsleading to abnormal activation of signal transduction pathways whichdrive malignant processes. Manning, G. et al., Science 2002, 298,1912-1934. Pharmacological inhibition of these signaling pathwayspresents promising intervention opportunities for targeted cancertherapies. Sawyers, C., Nature 2004, 432, 294-297.

Anaplastic lymphoma kinase (ALK), along with leukocyte tyrosine kinase(LTK), is grouped within the insulin receptor (IR) superfamily ofreceptor tyrosine kinases. ALK is mainly expressed in the central andperipheral nervous systems suggesting a potential role in normaldevelopment and function of the nervous system. Pulford, K. et al., CellMol. Life Sci. 2004, 61, 2939. ALK was first discovered as a fusionprotein, NPM (nucleophosmin)-ALK, encoded by a fusion gene arising fromthe t(2;5)(p23;q35) chromosomal translocation in anaplastic large celllymphoma (ALCL) cell lines. Morris, S. W. et al., Science 1994, 263,1281. More than twenty distinct ALK translocation partners have beendiscovered in many cancers, including ALCL (60-90% incidence),inflammatory myofibroblastic tumors (IMT, 50-60%), non-small cell lungcarcinomas (NSCLC, 3-7%), colorectal cancers (CRC, 0-2.4%), breastcancers (0-2.4%), and other carcinomas. Grande, E. et al., Mol. CancerTher. 2011, 10, 569-579. The ALK-fusion proteins are located in thecytoplasm, and the fusion partners with ALK play a role in dimerizationor oligomerization of the fusion proteins through a coil-coilinteraction to generate constitutive activation of ALK kinase function.Bischof, D. et al., Mol. Cell Biol., 1997, 17, 2312-2325. EML4-ALK,which comprises portions of the echinoderm microtubule associatedprotein-like 4 (EML4) gene and the ALK gene, was first discovered inNSCLC, is highly oncogenic, and was shown to cause lung adenocarcinomain transgenic mice. Soda, M. et al., Nature 2007, 448, 561-566.Oncogenic point mutations of ALK occur in both familial and sporadiccases of neuroblastoma. Mosse, Y. P. et al., Nature 2008, 455, 930-935.ALK is an attractive molecular target for cancer therapeuticintervention because of the important roles in haematopoietic, solid,and mesenchymal tumors. Grande, supra.

Since the first discovery of NPM (nucleophosmin)-ALK fusion gene inanaplastic large cell lymphoma cell lines in 1994 (Morris S W, et alScience. 1994; 263(5151):1281-4), alterations of ALK have been found ina wide range of cancer types, including anaplastic large cell lymphoma,inflammatory myofibroblastic tumor, diffuse large B cell lymphoma,NSCLC, renal medulla carcinoma, renal cell carcinoma, breast cancer,colon carcinoma, serous ovarian carcinoma, and esophageal squamous cellcarcinoma (Hallberg B, et al Nat Rev Cancer. 2013; 13(10):685-700). Theclinical benefit of targeting oncogenic ALK fusions over chemotherapy inALK+NSCLC patients has led to a full regular approval of crizotinib(Malik S M, et al Clin Cancer Res. 2014; 20(8):2029-34.). However,resistance to crizotinib treatment occurred within an average of7.3-10.9 months. The resistance mechanisms include ALK geneamplification, acquired mutations in the kinase domain, bypasssignaling, EMT, and CNS metastasis (Katayama R, et al Clin Cancer Res.2015; 21(10):2227-35). Although the more potent second generation ALKinhibitors ceritinib, alectinib and brigatinib can initially effectivelyovercome crizotinib resistance, patients ultimately relapse on theseinhibitors as well. Analysis of post-progression biopsy specimensindicated that each ALK inhibitor is associated with a distinct spectrumof ALK resistance mutations and ALK G1202R is a common resistancemutation to the first and second generations of ALK inhibitors (Gainor JF, et al Cancer Discov. 2016 October; 6(10):1118-1133). The solventfront mutations, including ALK G1202R, G1202del, D1203N, 51206Y/C, andE1210K increase significantly after second generation ALK inhibitortreatment, especially brigatinib having up to 71% solvent frontmutations in 7 biopsied resistant patients (Gainor J F, et al CancerDiscov. 2016 October; 6(10):1118-1133).

The current treatment paradigm for patients with ALK+cancer is to treatthem with sequential ALK targeted therapies. The third-generation ALKinhibitor lorlatinib has demonstrated clinical activity in patients whofailed previous ALK inhibitors and was approved for ALK positivemetastatic NSCLC patients after crizotinib and at least one other ALKinhibitor treatment (Shaw A, et al J Clin Oncol 2019, 37:1370-1379).However, Compound mutations emerge as a new challenge after more thanone ALK TKI treatment. Seven of 20 samples (35%) harbored compound ALKmutations after lorlatinib treatment, e. g. lorlatinib is no longeractive against compound gatekeeper and solvent front mutations ALKL1196M/G1202R (Yoda S., et al Cancer Discovery 2018, 8(6):714-729).

Endochondral ossification is a process that results in both thereplacement of the embryonic cartilaginous skeleton during organogenesisand the growth of long bones until adult height is achieved. Fibroblastgrowth factor (FGF)/FGF receptor (FGFR) signaling plays a vital role inthe development and maintenance of growth plates in endochondralossification process (Xie Y 2014). Missense mutations in FGFs and FGFRscan cause multiple genetic skeletal diseases with disorderedendochondral ossification. Activating mutations in FGFR3 causeachondroplasia, the most common form of dwarfism among live births(Samsa WE 2017). The growth plates of humans with FGFR3 mutations showdisrupted chondrocyte columns and reduced numbers of hypertrophicchondrocytes. FGFR1 and FGFR2 play many essential and mostly redundantroles during development, including growth plate formation.FGFR2-deficient embryos fail to form limb buds (Omitz D M 2015). Inaddition, Overexpression of FGFR1 in chondrocytes causes joint fusion.Deletion of both FGFR1 and FGFR2 in mice caused a decreased length ofthe growth plate with a reduced number of proliferating chondrocytes(Karuppaiah K 2016). Therefore, the selectivity over FGFRs is animportant parameter for better safety profile, especially for pediatricpopulation.

Therefore, there is a new demand to develop next generation ALKinhibitors that show potent activity against a broad spectrum ofresistant mutations, especially solvent front mutations, such as ALKG1202R, and compounds mutations, such as ALK L1196M/G1202R. Furthermore,there is a demand for such ALK inhibitors to show selectivity overFGFRs. Certain chiral diaryl macrocyclic compounds have been found inthe context of this disclosure to have this advantageous activityprofile.

SUMMARY

In one aspect, the disclosure relates to a compound of the formula

-   -   wherein    -   each R¹ and R² is independently H, deuterium, halogen, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic        heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),        —OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a),        —S(O)R^(a), —S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b),        —OS(O)NR^(a)R^(b), —OS(O)₂NR^(a)R^(b), —NR^(a)R^(b),        —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b),        —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b), —NR^(a)S(O)NR^(a)R^(b),        —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a), —C(O)OR^(a),        —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),        —P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b),        —P(O)OR^(a), —P(O)₂OR^(a), —CN, or —NO₂; or R¹ and R² taken        together with the carbon atom to which they are attached combine        to form a C₃-C₆ cycloalkyl; wherein each hydrogen atom in C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, and mono- or bicyclic        heteroaryl is independently optionally substituted by deuterium,        halogen, C₁-C₆ alkyl, C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   each R³ and R^(3′) is independently H, deuterium, halogen, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic        heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),        —OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a),        —S(O)R^(a), —S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b),        —OS(O)NR^(a)R^(b), —OS(O)₂NR^(a)R^(b), —NR^(a)R^(b),        —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b),        —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b), —NR^(a)S(O)NR^(a)R^(b),        —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a), —C(O)OR^(a),        —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),        —P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b),        —P(O)OR^(a), —P(O)₂OR^(a), —CN, or —NO₂, wherein each hydrogen        atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆        cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl,        mono- or bicyclic heteroaryl, is independently optionally        substituted by deuterium, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl,        —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f),        —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),        —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e),        —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f),        —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(e)R^(f),        —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f),        —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e), —C(O)OR^(e),        —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂; or R³ and R^(3′) taken        together with the carbon atom to which they are attached combine        to form a C₃-C₆ cycloalkyl; wherein each hydrogen atom in C₃-C₆        cycloalkyl is independently optionally substituted by deuterium,        halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂; or both R^(3′) taken        together combine to form a divalent group —(CR¹R²)_(m)—;    -   each R⁴ and R⁵ is independently hydrogen, deuterium, halogen,        —OR^(c), —OC(O)R^(c), —OC(O)NR^(c)R^(d), —OC(═N)NR^(c)R^(d),        —OS(O)R^(c), —OS(O)₂R^(c), —OS(O)NR^(c)R^(d),        —OS(O)₂NR^(c)R^(d), —SR^(c), —S(O)R^(c), —S(O)₂R^(c),        —S(O)NR^(c)R^(d), —S(O)₂NR^(c)R^(d), —NR^(c)R^(d),        —NR^(c)C(O)R^(d), —NR^(c)C(O)OR^(d), —NR^(c)C(O)NR^(c)R^(d),        —NR^(c)C(═N)NR^(c)R^(d), —NR^(c)S(O)R^(d), —NR^(c)S(O)₂R^(d),        —NR^(c)S(O)NR^(c)R^(d), —NR^(c)S(O)₂NR^(c)R^(d), —C(O)R^(c),        —C(O)OR^(c), —C(O)NR^(c)R^(d), —C(═N)NR^(c)R^(d), —PR^(c)R^(d),        —P(O)R^(c)R^(d), —P(O)₂R^(c)R^(d), —P(O)NR^(c)R^(d),        —P(O)₂NR^(c)R^(d), —P(O)OR^(c), —P(O)₂OR^(c), —CN, —NO₂, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic        heteroaryl, wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered        heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic heteroaryl,        C₅-C₈ cycloalkyl, or 5- to 8-membered heterocycloalkyl is        independently optionally substituted by deuterium, halogen,        C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   R⁶ is H, deuterium, or C₁-C₆ alkyl, wherein each hydrogen atom        in C₁-C₆ alkyl is independently optionally substituted by        deuterium, halogen, —OR^(e), —SR^(e), or —NR^(e)R^(f);    -   each R⁷ is independently hydrogen or deuterium;    -   each R⁸ and R⁹ is independently H, deuterium, halogen, —CN,        —OR^(e), C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆        cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, or        mono- or bicyclic heteroaryl, or alternatively, R⁸ and R⁹ taken        together with the carbon to which they are attached form a C₃-C₆        cycloalkyl or a 4- to 6-membered heterocycloalkyl, or        alternatively, R⁸ and R⁹ taken together with the carbon to which        they are attached form an exocyclic ethylene group, wherein each        hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,        C₃-C₆ cycloalkyl, 4- to 6-membered heterocycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, exocyclic ethylene        group, or mono- or bicyclic heteroaryl is optionally substituted        by a halogen, —N₃, —CN, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),        —OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e),        —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e),        —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f),        —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(e)R^(f),        —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f),        —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e), —C(O)OR^(e),        —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), or —P(O)₂OR^(e);    -   each R^(a), R^(b), R^(c), R^(d), R^(e), and R is independently        selected from the group consisting of H, deuterium, C₁-C₆ alkyl,        C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered        heterocycloalkyl, C₆-C₁₀ aryl, and 5- to 7-membered heteroaryl;    -   m is 1 or 2; and    -   n is 0, 1, 2, or 3;    -   or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure relates to a compound of the formula

-   -   wherein    -   each R¹ and R² is independently H, deuterium, halogen, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic        heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),        —OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a),        —S(O)R^(a), —S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b),        —OS(O)NR^(a)R^(b), —OS(O)₂NR^(a)R^(b), —NR^(a)R^(b),        —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b),        —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b), —NR^(a)S(O)NR^(a)R^(b),        —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a), —C(O)OR^(a),        —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),        —P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b),        —P(O)OR^(a), —P(O)₂OR^(a), —CN, or —NO₂; or R¹ and R² taken        together with the carbon atom to which they are attached combine        to form a C₃-C₆ cycloalkyl; wherein each hydrogen atom in C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, and mono- or bicyclic        heteroaryl is independently optionally substituted by deuterium,        halogen, C₁-C₆ alkyl, C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   each R³ and R^(3′) is independently H, deuterium, halogen, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic        heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),        —OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a),        —S(O)R^(a), —S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b),        —OS(O)NR^(a)R^(b), —OS(O)₂NR^(a)R^(b), —NR^(a)R^(b),        —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b),        —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b), —NR^(a)S(O)NR^(a)R^(b),        —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a), —C(O)OR^(a),        —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),        —P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b),        —P(O)OR^(a), —P(O)₂OR^(a), —CN, or —NO₂, wherein each hydrogen        atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆        cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl,        mono- or bicyclic heteroaryl, is independently optionally        substituted by deuterium, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl,        —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f),        —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),        —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e),        —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f),        —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(e)R^(f),        —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f),        —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e), —C(O)OR^(e),        —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂; or R³ and R^(3′) taken        together with the carbon atom to which they are attached combine        to form a C₃-C₆ cycloalkyl; wherein each hydrogen atom in C₃-C₆        cycloalkyl is independently optionally substituted by deuterium,        halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   each R⁴ and R⁵ is independently hydrogen, deuterium, halogen,        —OR^(c), —OC(O)R^(c), —OC(O)NR^(c)R^(d), —OC(═N)NR^(c)R^(d),        —OS(O)R^(c), —OS(O)₂R^(c), —OS(O)NR^(c)R^(d),        —OS(O)₂NR^(c)R^(d), —SR^(c), —S(O)R^(c), —S(O)₂R^(c),        —S(O)NR^(c)R^(d), —S(O)₂NR^(c)R^(d), —NR^(c)R^(d),        —NR^(c)C(O)R^(d), —NR^(c)C(O)OR^(d), —NR^(c)C(O)NR^(c)R^(d),        —NR^(c)C(═N)NR^(c)R^(d), —NR^(c)S(O)R^(d), —NR^(c)S(O)₂R^(d),        —NR^(c)S(O)NR^(c)R^(d), —NR^(c)S(O)₂NR^(c)R^(d), —C(O)R^(c),        —C(O)OR^(c), —C(O)NR^(c)R^(d), —C(═N)NR^(c)R^(d), —PR^(c)R^(d),        —P(O)R^(c)R^(d), —P(O)₂R^(c)R^(d), —P(O)NR^(c)R^(d),        —P(O)₂NR^(c)R^(d), —P(O)OR^(c), —P(O)₂OR^(c), —CN, —NO₂, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic        heteroaryl, wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered        heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic heteroaryl,        C₅-C₈ cycloalkyl, or 5- to 8-membered heterocycloalkyl is        independently optionally substituted by deuterium, halogen,        C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   R⁶ is H, deuterium, or C₁-C₆ alkyl, wherein each hydrogen atom        in C₁-C₆ alkyl is independently optionally substituted by        deuterium, halogen, —OR^(e), —SR^(e), or —NR^(e)R^(f);    -   each R⁷ is independently hydrogen or deuterium;    -   each R^(a), R^(b), R^(c), R^(d), R^(e), and R^(f) is        independently selected from the group consisting of H,        deuterium, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆        cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, and        5- to 7-membered heteroaryl; and    -   n is 0, 1, 2, or 3;    -   or a pharmaceutically acceptable salt thereof.

In an aspect of the compound of formula I or a pharmaceuticallyacceptable salt thereof:

-   -   each R¹ and R² is independently H or C₁-C₆ alkyl;    -   each R³ and R^(3′) is independently H or C₁-C₆ alkyl; or R³ and        R^(3′) taken together with the carbon atom to which they are        attached combine to form a C₃-C₆ cycloalkyl;    -   each R⁴ and R⁵ is independently hydrogen or halogen;    -   R⁶ is H or C₁-C₆ alkyl;    -   each R⁷ is hydrogen; and    -   n is 0 or 1.

In an aspect of the compound of formula I or a pharmaceuticallyacceptable salt thereof:

-   -   each R¹ and R² is independently H or C₁-C₃ alkyl;    -   each R³ and R^(3′) is independently H or C₁-C₃ alkyl; or R³ and        R^(3′) taken together with the carbon atom to which they are        attached combine to form a C₃-C₄ cycloalkyl;    -   each R⁴ and R⁵ is independently hydrogen or halogen;    -   R⁶ is H;    -   each R⁷ is hydrogen; and    -   n is 0 or 1.

In an aspect of the compound of formula I or a pharmaceuticallyacceptable salt thereof.

-   -   each R¹ and R² is independently hydrogen or methyl;    -   each R³ and R^(3′) is independently hydrogen or methyl; or R³        and R^(3′) taken together with the carbon atom to which they are        attached combine to form a cyclopropyl;    -   each R⁴ and R⁵ is independently hydrogen or fluorine;    -   R⁶ is hydrogen;    -   each R⁷ is hydrogen; and    -   n is 0 or 1.

In an aspect of the compound of formula I, or a pharmaceuticallyacceptable salt thereof, has the structure of formula II, IIa, III, orIIIa.

In another aspect, the disclosure relates to a compound of the formulaII

-   -   or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure relates to a compound of the formulaIIa

-   -   or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure relates to a compound of the formulaIII

-   -   or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure relates to a compound of the formulaIIIa

-   -   or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure provides a pharmaceutical compositioncomprising a compound of any one of the disclosed aspects, or apharmaceutically acceptable salt thereof, and optionally at least onediluent, carrier or excipient.

In another aspect, the disclosure provides a method of treating cancercomprising administering to a subject in need of such treatment aneffective amount of at least one compound of any one of the disclosedaspects, or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure provides a use of a compound of anyone of the disclosed aspects, or a pharmaceutically acceptable saltthereof, in the preparation of a medicament for the treatment of cancer.

In another aspect, the disclosure provides a use of a compound of anyone of the disclosed aspects, or a pharmaceutically acceptable saltthereof, for treating cancer.

In another aspect, the disclosure provides a compound of the disclosedaspects, or a pharmaceutically acceptable salt thereof, for use intreating cancer in a subject.

In another aspect, the disclosure provides a method of inhibiting ALKreceptor tyrosine kinase, comprising contacting a cell comprising one ormore of such kinases with an effective amount of at least one compoundof any one of the disclosed aspects, or a pharmaceutically acceptablesalt thereof, and/or with at least one pharmaceutical composition of thedisclosure, wherein the contacting is in vitro, ex vivo, or in vivo.

Additional embodiments, features, and advantages of the disclosure willbe apparent from the following detailed description and through practiceof the disclosure. The compounds of the present disclosure can bedescribed as embodiments in any of the following enumerated clauses. Itwill be understood that any of the embodiments described herein can beused in connection with any other embodiments described herein to theextent that the embodiments do not contradict one another.

1. A compound of the formula I

-   -   wherein    -   each R¹ and R² is independently H, deuterium, halogen, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic        heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),        —OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a),        —S(O)R^(a), —S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b),        —OS(O)NR^(a)R^(b), —OS(O)₂NR^(a)R^(b), —NR^(a)R^(b),        —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b),        —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b), —NR^(a)S(O)NR^(a)R^(b),        —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a), —C(O)OR^(a),        —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),        —P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b),        —P(O)OR^(a), —P(O)₂OR^(a), —CN, or —NO₂; or R¹ and R² taken        together with the carbon atom to which they are attached combine        to form a C₃-C₆ cycloalkyl; wherein each hydrogen atom in C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, and mono- or bicyclic        heteroaryl is independently optionally substituted by deuterium,        halogen, C₁-C₆ alkyl, C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   each R³ and R^(3′) is independently H, deuterium, halogen, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic        heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),        —OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a),        —S(O)R^(a), —S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b),        —OS(O)NR^(a)R^(b), —OS(O)₂NR^(a)R^(b), —NR^(a)R^(b),        —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b),        —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b), —NR^(a)S(O)NR^(a)R^(b),        —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a), —C(O)OR^(a),        —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),        —P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b),        —P(O)OR^(a), —P(O)₂OR^(a), —CN, or —NO₂, wherein each hydrogen        atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆        cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl,        mono- or bicyclic heteroaryl, is independently optionally        substituted by deuterium, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl,        —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f),        —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),        —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e),        —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f),        —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(e)R^(f),        —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f),        —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e), —C(O)OR^(e),        —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂; or R³ and R^(3′) taken        together with the carbon atom to which they are attached combine        to form a C₃-C₆ cycloalkyl; wherein each hydrogen atom in C₃-C₆        cycloalkyl is independently optionally substituted by deuterium,        halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   each R⁴ and R⁵ is independently hydrogen, deuterium, halogen,        —OR^(c), —OC(O)R^(c), —OC(O)NR^(c)R^(d), —OC(═N)NR^(c)R^(d),        —OS(O)R^(c), —OS(O)₂R^(c), —OS(O)NR^(c)R^(d),        —OS(O)₂NR^(c)R^(d), —SR^(c), —S(O)R^(c), —S(O)₂R^(c),        —S(O)NR^(c)R^(d), —S(O)₂NR^(c)R^(d), —NR^(c)R^(d),        —NR^(c)C(O)R^(d), —NR^(c)C(O)OR^(d), —NR^(c)C(O)NR^(c)R^(d),        —NR^(e)C(═N)NR^(c)R^(d), —NR^(c)S(O)R^(d), —NR^(c)S(O)₂R^(d),        —NR^(c)S(O)NR^(c)R^(d), —NR^(c)S(O)₂NR^(c)R^(d), —C(O)R^(c),        —C(O)OR^(c), —C(O)NR^(c)R^(d), —C(═N)NR^(c)R^(d), —PR^(c)R^(d),        —P(O)R^(c)R^(d), —P(O)₂R^(c)R^(d), —P(O)NR^(c)R^(d),        —P(O)₂NR^(c)R^(d), —P(O)OR^(c), —P(O)₂OR^(c), —CN, —NO₂, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to        7-membered heterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic        heteroaryl, wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered        heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic heteroaryl,        C₅-C₈cycloalkyl, or 5- to 8-membered heterocycloalkyl is        independently optionally substituted by deuterium, halogen,        C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),        —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e),        —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e),        —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),        —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),        —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),        —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),        —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),        —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),        —P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂;    -   R⁶ is H, deuterium, or C₁-C₆ alkyl, wherein each hydrogen atom        in C₁-C₆ alkyl is independently optionally substituted by        deuterium, halogen, —OR^(e), —SR^(e), or —NR^(e)R^(f);    -   each R⁷ is independently hydrogen or deuterium;    -   each R^(a), R^(b), R^(c), R^(d), R^(e), and R^(f) is        independently selected from the group consisting of H,        deuterium, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆        cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, and        5- to 7-membered heteroaryl; and    -   n is 0, 1, 2, or 3;    -   or a pharmaceutically acceptable salt thereof.

2. The compound of clause 1, or a pharmaceutically acceptable saltthereof, wherein n is 0 or 1.

3. The compound of clause 1 or 2, or a pharmaceutically acceptable saltthereof, wherein n is 0.

4. The compound of clause 1 or 2, or a pharmaceutically acceptable saltthereof, wherein n is 1.

5. The compound of any one of clauses 1 to 3, having the formula II

-   -   or a pharmaceutically acceptable salt thereof.

6. The compound of any one of clauses 1 to 3, or 5, having the formulaIIa

-   -   or a pharmaceutically acceptable salt thereof.

7. The compound of any one of clauses 1, 2, or 4, having the formula III

-   -   or a pharmaceutically acceptable salt thereof.

8. The compound of any one of clauses 1, 2, 4, or 7, having the formulaIIIa

-   -   or a pharmaceutically acceptable salt thereof.

9. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein each R¹ and R², whenpresent, is independently H, deuterium, C₁-C₆ alkyl, or R¹ and R² takentogether with the carbon atom to which they are attached combine to forma C₃-C₆ cycloalkyl.

10. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein each R³ and R^(3′) isindependently H, deuterium, C₁-C₆ alkyl, or R³ and R^(3′) taken togetherwith the carbon atom to which they are attached combine to form a C₃-C₆cycloalkyl.

11. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein each R^(3′) is H ordeuterium.

12. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein each R⁴ is H ordeuterium.

13. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein R⁵ is fluoro.

14. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein R⁶ is H.

15. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein each R⁷ is H.

16. The compound of clause 1, selected from the group consisting of

-   -   or a pharmaceutically acceptable salt thereof.

17. A pharmaceutical composition comprising a compound of any one of thepreceding clauses, or a pharmaceutically acceptable salt thereof, andoptionally at least one diluent, carrier or excipient.

18. A method of treating cancer, pain, neurological diseases, autoimmunediseases, or inflammation comprising administering to a subject in needof such treatment an effective amount of at least one compound of anyone of clauses 1 to 16, or a pharmaceutically acceptable salt thereof.

19. The method of clause 18, wherein the disease is cancer.

20. The method of clause 18 or 19, wherein the subject is a human.

21. The method of any one of clauses 18 to 20, wherein the disease is acancer mediated by ALK.

22. The method of any one of clauses 18 to 21, wherein the disease is acancer mediated by a genetically altered ALK.

23. The method of any one of clauses 18 to 22, wherein the disease is acancer mediated by a fusion protein comprising a fragment of a proteinencoded by an ALK gene and a fragment of a protein encoded by a geneselected from the group consisting of NPM, EML4, TPR, TFG, ATIC, CLTC1,TPM4, MSN ALO17 and MYH9.

24. The method of clause 23, wherein the fusion protein comprises afragment of a protein encoded by an ALK gene and a fragment of a proteinencoded by an EML4 gene.

25. The method of clause 23, wherein the genetically altered ALK is anEML4-ALK fusion protein.

26. The method of clause 25, wherein the EML4-ALK fusion protein is awild-type protein.

27. The method of clause 25, wherein the EML4-ALK fusion proteincomprises at least one resistance mutation.

28. The method of clause 25, wherein the EML4-ALK fusion proteincomprises at least one mutation selected from the group consisting of toL1196M, G1202R, C1156Y, D1203N, G1202 deletion, E1210K, S1206C, F1174C,F1174L, F1174S, F1174V, F1245C, G1269A, G1269S, I1171N, L1152P, L1152R,L1198F, R1275Q, S1206R, T1151-L1152insT, T1151M, V1180L, andcombinations thereof.

29. The method of clause 25, wherein the EML4-ALK fusion proteincomprises a mutation combination selected from the group consisting ofE1210K/D1203N, E1210K/S1206C, L1198F/C1156Y, L1198F/G1202R,L1198F/L1196M, L1196M/G1202R, L1198F/C1156Y, G1202R/G1269A, andG1202R/G1269A/L1204V.

30. The method of any one of clauses 18 to 29, wherein the disease is acancer selected from the group consisting of ALCL, NSCLC, neuroblastoma,inflammatory myofibroblastic tumor, adult renal cell carcinoma,pediatric renal cell carcinoma, breast cancer, ER⁺ breast cancer,colonic adenocarcinoma, glioblastoma, glioblastoma multiforme,anaplastic thyroid cancer, cholangiocarcinoma, ovarian cancer, gastricadenocarcinoma, colorectal cancer, inflammatory myofibroblastic tumor,angiosarcoma, epithelioid hemangioendothelioma, intrahepaticcholangiocarcinoma, thyroid papillary cancer, spitzoid neoplasms,sarcoma, astrocytoma, brain lower grade glioma, secretory breastcarcinoma, mammary analogue carcinoma, acute myeloid leukemia,congenital mesoblastic nephroma, congenital fibrosarcomas, Ph-like acutelymphoblastic leukemia, thyroid carcinoma, skin cutaneous melanoma, headand neck squamous cell carcinoma, pediatric glioma CML, prostate cancer,lung squamous carcinoma, ovarian serous cystadenocarcinoma, skincutaneous melanoma, castrate-resistant prostate cancer, Hodgkinlymphoma, and serous and clear cell endometrial cancer.

31. Use of a compound of any one of clauses 1 to 16, or apharmaceutically acceptable salt thereof, in the preparation of amedicament for the treatment of cancer.

32. Use of a compound of any one of clauses 1 to 16, or apharmaceutically acceptable salt thereof, for treating cancer.

33. The use of clause 31 or 32, wherein the disease is a cancer mediatedby ALK.

34. The use of any one of clauses 31 to 33, wherein the disease is acancer mediated by a genetically altered ALK.

35. The use of any one of clauses 31 to 35, wherein the disease is acancer mediated by a fusion protein comprising a fragment of a proteinencoded by an ALK gene and a fragment of a protein encoded by a geneselected from the group consisting of NPM, EML4, TPR, TFG, ATIC, CLTC1,TPM4, MSN ALO17 and MYH9.

36. The use of clause 35, wherein the fusion protein comprises afragment of a protein encoded by an ALK gene and a fragment of a proteinencoded by an EML4 gene.

37. The use of clause 35, wherein the genetically altered ALK is anEML4-ALK fusion protein.

38. The use of clause 37, wherein the EML4-ALK fusion protein is awild-type protein.

39. The use of clause 37, wherein the EML4-ALK fusion protein comprisesat least one resistance mutation.

40. The use of clause 37, wherein the EML4-ALK fusion protein comprisesat least one mutation selected from the group consisting of to L1196M,G1202R, C1156Y, D1203N, G1202 deletion, E1210K, S1206C, F1174C, F1174L,F1174S, F1174V, F1245C, G1269A, G1269S, I1171N, L1152P, L1152R, L1198F,R1275Q, S1206R, T1151-L1152insT, T1151M, V1180L, and combinationsthereof.

41. The use of clause 37, wherein the EML4-ALK fusion protein comprisesa mutation combination selected from the group consisting ofE1210K/D1203N, E1210K/S1206C, L1198F/C₁₁₅₆Y, L1198F/G1202R,L1198F/L1196M, L1196M/G1202R, L1198F/C₁₁₅₆Y, G1202R/G1269A, andG1202R/G1269A/L1204V.

42. The use of any one of clauses 31 to 41, wherein the cancer selectedfrom the group consisting of ALCL, NSCLC, neuroblastoma, inflammatorymyofibroblastic tumor, adult renal cell carcinoma, pediatric renal cellcarcinoma, breast cancer, ER⁺ breast cancer, colonic adenocarcinoma,glioblastoma, glioblastoma multiforme, anaplastic thyroid cancer,cholangiocarcinoma, ovarian cancer, gastric adenocarcinoma, colorectalcancer, inflammatory myofibroblastic tumor, angiosarcoma, epithelioidhemangioendothelioma, intrahepatic cholangiocarcinoma, thyroid papillarycancer, spitzoid neoplasms, sarcoma, astrocytoma, brain lower gradeglioma, secretory breast carcinoma, mammary analogue carcinoma, acutemyeloid leukemia, congenital mesoblastic nephroma, congenitalfibrosarcomas, Ph-like acute lymphoblastic leukemia, thyroid carcinoma,skin cutaneous melanoma, head and neck squamous cell carcinoma,pediatric glioma CML, prostate cancer, lung squamous carcinoma, ovarianserous cystadenocarcinoma, skin cutaneous melanoma, castrate-resistantprostate cancer, Hodgkin lymphoma, and serous and clear cell endometrialcancer.

43. A method of inhibiting ALK receptor tyrosine kinase, comprisingcontacting a cell comprising one or more of such kinases with aneffective amount of a compound of any one of clauses 1 to 16, or apharmaceutically acceptable salt thereof, and/or with at least onepharmaceutical composition comprising a compound of any one of clauses 1to 16, wherein the contacting is in vitro, ex vivo, or in vivo.

44. A compound of any one of clauses 1 to 16, for use in treating cancerin a subject.

45. The compound of clause 44, wherein the disease is a cancer mediatedby ALK.

46. The compound of clause 44 or 45, wherein the disease is a cancermediated by a genetically altered ALK.

47. The compound of any one of clauses 44 to 46, wherein the disease isa cancer mediated by a fusion protein comprising a fragment of a proteinencoded by an ALK gene and a fragment of a protein encoded by a geneselected from the group consisting of NPM, EML4, TPR, TFG, ATIC, CLTC1,TPM4, MSN ALO17 and MYH9.

48. The compound of clause 47, wherein the fusion protein comprises afragment of a protein encoded by an ALK gene and a fragment of a proteinencoded by an EML4 gene.

49. The compound of clause 47, wherein the genetically altered ALK is anEML4-ALK fusion protein.

50. The compound of clause 49, wherein the EML4-ALK fusion protein is awild-type protein.

51. The compound of clause 49, wherein the EML4-ALK fusion proteincomprises at least one resistance mutation.

52. The compound of clause 49, wherein the EML4-ALK fusion proteincomprises at least one mutation selected from the group consisting of toL1196M, G1202R, C1156Y, D1203N, G1202 deletion, E1210K, S1206C, F1174C,F1174L, F1174S, F1174V, F1245C, G1269A, G1269S, I1171N, L1152P, L1152R,L1198F, R1275Q, S1206R, T1151-L1152insT, T1151M, V1180L, andcombinations thereof.

53. The compound of clause 49, wherein the EML4-ALK fusion proteincomprises a mutation combination selected from the group consisting ofE1210K/D1203N, E1210K/S1206C, L1198F/C1156Y, L1198F/G1202R,L1198F/L1196M, L1196M/G1202R, L1198F/C1156Y, G1202R/G1269A, andG1202R/G1269A/L1204V.

54. The compound of any one of clauses 44 to 53, wherein the cancerselected from the group consisting of ALCL, NSCLC, neuroblastoma,inflammatory myofibroblastic tumor, adult renal cell carcinoma,pediatric renal cell carcinoma, breast cancer, ER breast cancer, colonicadenocarcinoma, glioblastoma, glioblastoma multiforme, anaplasticthyroid cancer, cholangiocarcinoma, ovarian cancer, gastricadenocarcinoma, colorectal cancer, inflammatory myofibroblastic tumor,angiosarcoma, epithelioid hemangioendothelioma, intrahepaticcholangiocarcinoma, thyroid papillary cancer, spitzoid neoplasms,sarcoma, astrocytoma, brain lower grade glioma, secretory breastcarcinoma, mammary analogue carcinoma, acute myeloid leukemia,congenital mesoblastic nephroma, congenital fibrosarcomas, Ph-like acutelymphoblastic leukemia, thyroid carcinoma, skin cutaneous melanoma, headand neck squamous cell carcinoma, pediatric glioma CML, prostate cancer,lung squamous carcinoma, ovarian serous cystadenocarcinoma, skincutaneous melanoma, castrate-resistant prostate cancer, Hodgkinlymphoma, and serous and clear cell endometrial cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows tumor volumes of xenograft tumor models in mice bearingBa/F3 cell derived xenograft tumors harboring an EML4-ALK fusion with aG1202R mutation. (●) vehicle; (▪) Compound 1 (2 mg/kg BID); (▴) Compound1 (5 mg/kg BID); (▾) Compound 1 (10 mg/kg BID); (

) Compound 2 (3 mg/kg BID); (Δ) Compound 2 (10 mg/kg BID); (♦)Lorlatinib (5 mg/kg).

FIG. 1B shows body weights of mice bearing Ba/F3 cell derived xenografttumors harboring an EML4-ALK fusion with a G1202R mutation. (●) vehicle;(▪) Compound 1 (2 mg/kg BID); (▴) Compound 1 (5 mg/kg BID); (▾) Compound1 (10 mg/kg BID); (

) Compound 2 (3 mg/kg BID); (Δ) Compound 2 (10 mg/kg BID); (♦)Lorlatinib (5 mg/kg).

FIG. 2A shows tumor volumes of xenograft tumor models in mice bearingBa/F3 cell derived xenograft tumors harboring an EML4-ALK fusion with aL1198F/G1202R mutation. (●) vehicle; (▪) Compound 1 (2 mg/kg BID); (▴)Compound 1 (5 mg/kg BID); (▾) Compound 1 (10 mg/kg BID); (♦) Lorlatinib(5 mg/kg).

FIG. 2B shows body weights of mice bearing Ba/F3 cell derived xenografttumors harboring an EML4-ALK fusion with a L1198F/G1202R mutation. (●)vehicle; (▪) Compound 1 (2 mg/kg BID); (▴) Compound 1 (5 mg/kg BID); (▾)Compound 1 (10 mg/kg BID); (♦) Lorlatinib (5 mg/kg).

FIG. 3A shows tumor volumes of xenograft tumor models in mice bearingBa/F3 cell derived xenograft tumors harboring an EML4-ALK fusion with aL1196M/G1202R mutation. (●) vehicle; (

) Compound 1 (0.6 mg/kg BID); (▪) Compound 1 (2 mg/kg BID); (▴) Compound1 (5 mg/kg BID); (▾) Compound 1 (10 mg/kg BID); (♦) Lorlatinib (5mg/kg).

FIG. 3B shows body weights of mice bearing Ba/F3 cell derived xenografttumors harboring an EML4-ALK fusion with a L1196M/G1202R mutation. (●)vehicle; (

) Compound 1 (0.6 mg/kg BID); (▪) Compound 1 (2 mg/kg BID); (▴) Compound1 (5 mg/kg BID); (▾) Compound 1 (10 mg/kg BID); (♦) Lorlatinib (5mg/kg).

FIG. 4 shows the inhibition of phosphorylation of ALK fusions atY1282/1283 and at Y1604, and reduction in the level of ALK fusionexpression in mice treated with Compound 1 at 10 mg/kg BID. As thecontrol, the expression level of actin was not affected by Compound 1treatment.

FIG. 5 shows the inhibition of phosphorylation of ALK fusions as afunction of free plasma concentration of compound upon dosing ofCompound 1 at 0.6, 2, or 5 mg/kg after 2 hours or 12 hours in micebearing Ba/F3 cell derived xenograft tumors harboring an EML4-ALK fusionwith a L1196M/G1202R mutation.

DETAILED DESCRIPTION

Before the present disclosure is further described, it is to beunderstood that this disclosure is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present disclosure will be limited only by the appendedclaims.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art to which this disclosure belongs. All patents, applications,published applications and other publications referred to herein areincorporated by reference in their entireties. If a definition set forthin this section is contrary to or otherwise inconsistent with adefinition set forth in a patent, application, or other publication thatis herein incorporated by reference, the definition set forth in thissection prevails over the definition incorporated herein by reference.

As used herein and in the appended claims, the singular forms “a,” “an,”and “the” include plural referents unless the context clearly dictatesotherwise. It is further noted that the claims may be drafted to excludeany optional element. As such, this statement is intended to serve asantecedent basis for use of such exclusive terminology as “solely,”“only” and the like in connection with the recitation of claim elements,or use of a “negative” limitation.

As used herein, the terms “including,” “containing,” and “comprising”are used in their open, non-limiting sense.

To provide a more concise description, some of the quantitativeexpressions given herein are not qualified with the term “about.” It isunderstood that, whether the term “about” is used explicitly or not,every quantity given herein is meant to refer to the actual given value,and it is also meant to refer to the approximation to such given valuethat would reasonably be inferred based on the ordinary skill in theart, including equivalents and approximations due to the experimentaland/or measurement conditions for such given value. Whenever a yield isgiven as a percentage, such yield refers to a mass of the entity forwhich the yield is given with respect to the maximum amount of the sameentity that could be obtained under the particular stoichiometricconditions. Concentrations that are given as percentages refer to massratios, unless indicated differently.

Except as otherwise noted, the methods and techniques of the presentembodiments are generally performed according to conventional methodswell known in the art and as described in various general and morespecific references that are cited and discussed throughout the presentspecification. See, e.g., Loudon, Organic Chemistry, Fourth Edition, NewYork: Oxford University Press, 2002, pp. 360-361, 1084-1085; Smith andMarch, March's Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, Fifth Edition, Wiley-Interscience, 2001. Chemicalnomenclature for compounds described herein has generally been derivedusing the commercially-available ACD/Name 2014 (ACD/Labs) or ChemBioDrawUltra 13.0 (Perkin Elmer).

It is appreciated that certain features of the disclosure, which are,for clarity, described in the context of separate embodiments, may alsobe provided in combination in a single embodiment. Conversely, variousfeatures of the disclosure, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination. All combinations of the embodimentspertaining to the chemical groups represented by the variables arespecifically embraced by the present disclosure and are disclosed hereinjust as if each and every combination was individually and explicitlydisclosed, to the extent that such combinations embrace compounds thatare stable compounds (i.e., compounds that can be isolated,characterized, and tested for biological activity). In addition, allsubcombinations of the chemical groups listed in the embodimentsdescribing such variables are also specifically embraced by the presentdisclosure and are disclosed herein just as if each and every suchsub-combination of chemical groups was individually and explicitlydisclosed herein.

Definitions

As used herein, the term “alkyl” includes a chain of carbon atoms, whichis optionally branched and contains from 1 to 20 carbon atoms. It is tobe further understood that in certain embodiments, alkyl may beadvantageously of limited length, including C₁-C₁₂, C₁-C₁₀, C₁-C₉,C₁-C₈, C₁-C₇, C₁-C₆, and C₁-C₄, Illustratively, such particularlylimited length alkyl groups, including C₁-C₈, C₁-C₇, C₁-C₆, and C₁-C₄,and the like may be referred to as “lower alkyl.” Illustrative alkylgroups include, but are not limited to, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, 2-pentyl,3-pentyl, neopentyl, hexyl, heptyl, octyl, and the like. Alkyl may besubstituted or unsubstituted. Typical substituent groups includecycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy,mercapto, alkylthio, arylthio, cyano, halo, carbonyl, oxo, (═O),thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, nitro, and amino, or asdescribed in the various embodiments provided herein. It will beunderstood that “alkyl” may be combined with other groups, such as thoseprovided above, to form a functionalized alkyl. By way of example, thecombination of an “alkyl” group, as described herein, with a “carboxy”group may be referred to as a “carboxyalkyl” group. Other non-limitingexamples include hydroxyalkyl, aminoalkyl, and the like.

As used herein, the term “alkenyl” includes a chain of carbon atoms,which is optionally branched, and contains from 2 to 20 carbon atoms,and also includes at least one carbon-carbon double bond (i.e., C═C). Itwill be understood that in certain embodiments, alkenyl may beadvantageously of limited length, including C₂-C₁₂, C₂-C₉, C₂-C₈, C₂-C₇,C₂-C₆, and C₂-C₄. Illustratively, such particularly limited lengthalkenyl groups, including C₂-C₈, C₂-C₇, C₂-C₆, and C₂-C₄ may be referredto as lower alkenyl. Alkenyl may be unsubstituted, or substituted asdescribed for alkyl or as described in the various embodiments providedherein. Illustrative alkenyl groups include, but are not limited to,ethenyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, and the like.

As used herein, the term “alkynyl” includes a chain of carbon atoms,which is optionally branched, and contains from 2 to 20 carbon atoms,and also includes at least one carbon-carbon triple bond (i.e., C≡C). Itwill be understood that in certain embodiments, alkynyl may each beadvantageously of limited length, including C₂-C₁₂, C₂-C₉, C₂-C₈, C₂-C₇,C₂-C₆, and C₂-C₄. Illustratively, such particularly limited lengthalkynyl groups, including C₂-C₈, C₂-C₇, C₂-C₆, and C₂-C₄ may be referredto as lower alkynyl. Alkynyl may be unsubstituted, or substituted asdescribed for alkyl or as described in the various embodiments providedherein. Illustrative alkynyl groups include, but are not limited to,ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, and the like.

As used herein, the term “aryl” refers to an all-carbon monocyclic orfused-ring polycyclic groups of 6 to 12 carbon atoms having a completelyconjugated pi-electron system. It will be understood that in certainembodiments, aryl may be advantageously of limited size such as C₆-C₁₀aryl. Illustrative aryl groups include, but are not limited to, phenyl,naphthylenyl and anthracenyl. The aryl group may be unsubstituted, orsubstituted as described for alkyl or as described in the variousembodiments provided herein.

As used herein, the term “cycloalkyl” refers to a 3 to 15 memberall-carbon monocyclic ring, including an all-carbon 5-member/6-member or6-member/6-member fused bicyclic ring, or a multicyclic fused ring (a“fused” ring system means that each ring in the system shares anadjacent pair of carbon atoms with each other ring in the system) group,or a carbocyclic ring that is fused to another group such as aheterocyclic, such as ring 5- or 6-membered cycloalkyl fused to a 5- to7-membered heterocyclic ring, where one or more of the rings may containone or more double bonds but the cycloalkyl does not contain acompletely conjugated pi-electron system. It will be understood that incertain embodiments, cycloalkyl may be advantageously of limited sizesuch as C₃-C₁₃, C₃-C₉, C₃-C₆ and C₄-C₆. Cycloalkyl may be unsubstituted,or substituted as described for alkyl or as described in the variousembodiments provided herein. Illustrative cycloalkyl groups include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl,cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl, adamantyl,norbonyl, norbornenyl, 9H-fluoren-9-yl, and the like. Illustrativeexamples of cycloalkyl groups shown in graphical representations includethe following entities, in the form of properly bonded moieties:

As used herein, the term “heterocycloalkyl” refers to a monocyclic orfused ring group having in the ring(s) from 3 to 12 ring atoms, in whichat least one ring atom is a heteroatom, such as nitrogen, oxygen orsulfur, the remaining ring atoms being carbon atoms. Heterocycloalkylmay optionally contain 1, 2, 3 or 4 heteroatoms. A heterocycloalkylgroup may be fused to another group such as another heterocycloalkyl, ora heteroaryl group. Heterocycloalkyl may also have one of more doublebonds, including double bonds to nitrogen (e.g., C═N or N═N) but doesnot contain a completely conjugated pi-electron system. It will beunderstood that in certain embodiments, heterocycloalkyl may beadvantageously of limited size such as 3- to 7-memberedheterocycloalkyl, 5- to 7-membered heterocycloalkyl, 3-, 4-, 5- or6-membered heterocycloalkyl, and the like. Heterocycloalkyl may beunsubstituted, or substituted as described for alkyl or as described inthe various embodiments provided herein. Illustrative heterocycloalkylgroups include, but are not limited to, oxiranyl, thianaryl, azetidinyl,oxetanyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl,piperidinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl, piperazinyl,oxepanyl, 3,4-dihydro-2H-pyranyl, 5,6-dihydro-2H-pyranyl, 2H-pyranyl, 1,2, 3, 4-tetrahydropyridinyl, and the like. Illustrative examples ofheterocycloalkyl groups shown in graphical representations include thefollowing entities, in the form of properly bonded moieties:

As used herein, the term “heteroaryl” refers to a monocyclic or fusedring group of 5 to 12 ring atoms containing one, two, three or four ringheteroatoms selected from nitrogen, oxygen and sulfur, the remainingring atoms being carbon atoms, and also having a completely conjugatedpi-electron system. It will be understood that in certain embodiments,heteroaryl may be advantageously of limited size such as 3- to7-membered heteroaryl, 5- to 7-membered heteroaryl, and the like.Heteroaryl may be unsubstituted, or substituted as described for alkylor as described in the various embodiments provided herein. Illustrativeheteroaryl groups include, but are not limited to, pyrrolyl, furanyl,thiophenyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl,pyrimidinyl, quinolinyl, isoquinolinyl, purinyl, tetrazolyl, triazinyl,pyrazinyl, tetrazinyl, quinazolinyl, quinoxalinyl, thienyl, isoxazolyl,isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzimidazolyl,benzoxazolyl, benzthiazolyl, benzisoxazolyl, benzisothiazolyl andcarbazoloyl, and the like. Illustrative examples of heteroaryl groupsshown in graphical representations, include the following entities, inthe form of properly bonded moieties:

As used herein, “hydroxy” or “hydroxyl” refers to an —OH group.

As used herein, “alkoxy” refers to both an —O-(alkyl) or an—O-(unsubstituted cycloalkyl) group. Representative examples include,but are not limited to, methoxy, ethoxy, propoxy, butoxy,cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and thelike.

As used herein, “aryloxy” refers to an —O-aryl or an —O-heteroarylgroup. Representative examples include, but are not limited to, phenoxy,pyridinyloxy, furanyloxy, thienyloxy, pyrimidinyloxy, pyrazinyloxy, andthe like, and the like.

As used herein, “mercapto” refers to an —SH group.

As used herein, “alkylthio” refers to an —S-(alkyl) or an—S-(unsubstituted cycloalkyl) group. Representative examples include,but are not limited to, methylthio, ethylthio, propylthio, butylthio,cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio, andthe like.

As used herein, “arylthio” refers to an —S-aryl or an —S-heteroarylgroup. Representative examples include, but are not limited to,phenylthio, pyridinylthio, furanylthio, thienylthio, pyrimidinylthio,and the like.

As used herein, “halo” or “halogen” refers to fluorine, chlorine,bromine or iodine.

As used herein, “cyano” refers to a —CN group.

The term “oxo” represents a carbonyl oxygen. For example, a cyclopentylsubstituted with oxo is cyclopentanone.

As used herein, “bond” refers to a covalent bond.

The term “substituted” means that the specified group or moiety bearsone or more substituents. The term “unsubstituted” means that thespecified group bears no substituents. Where the term “substituted” isused to describe a structural system, the substitution is meant to occurat any valency-allowed position on the system. In some embodiments,“substituted” means that the specified group or moiety bears one, two,or three substituents. In other embodiments, “substituted” means thatthe specified group or moiety bears one or two substituents. In stillother embodiments, “substituted” means the specified group or moietybears one substituent.

As used herein, “optional” or “optionally” means that the subsequentlydescribed event or circumstance may but need not occur, and that thedescription includes instances where the event or circumstance occursand instances in which it does not. For example, “wherein each hydrogenatom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3-to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclicheteroaryl is independently optionally substituted by C₁-C₆ alkyl” meansthat an alkyl may be but need not be present on any of the C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic heteroaryl byreplacement of a hydrogen atom for each alkyl group, and the descriptionincludes situations where the C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, ormono- or bicyclic heteroaryl is substituted with an alkyl group andsituations where the C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, or mono- orbicyclic heteroaryl is not substituted with the alkyl group.

As used herein, “independently” means that the subsequently describedevent or circumstance is to be read on its own relative to other similarevents or circumstances. For example, in a circumstance where severalequivalent hydrogen groups are optionally substituted by another groupdescribed in the circumstance, the use of “independently optionally”means that each instance of a hydrogen atom on the group may besubstituted by another group, where the groups replacing each of thehydrogen atoms may be the same or different. Or for example, wheremultiple groups exist all of which can be selected from a set ofpossibilities, the use of “independently” means that each of the groupscan be selected from the set of possibilities separate from any othergroup, and the groups selected in the circumstance may be the same ordifferent.

As used herein, the phrase “taken together with the carbon atom to whichthey are attached” or “taken together with the carbon atom to which theyare attached” means that two substituents (e.g., R¹ and R²) attached tothe same carbon atom form the groups that are defined by the claim, suchas C₃-C₆ cycloalkyl. In particular, the phrase “taken together with thecarbon atom to which they are attached” means that when, for example, R¹and R², and the carbon atom to which they are attached form a C₃-C₆cycloalkyl, then the formed ring will be attached at the same carbonatom. For example, the phrase “R¹ and R² taken together with the carbonatom to which they are attached form a C₃-C₆ cycloalkyl” used inconnection with the embodiments described herein includes the compoundsrepresented as follows:

where the above spirocyclic rings can be optionally substituted asdefined in the given embodiment.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which counter ions which may be used in pharmaceuticals.See, generally, S. M. Berge, et al., “Pharmaceutical Salts,” J. Pharm.Sci., 1977, 66, 1-19. Preferred pharmaceutically acceptable salts arethose that are pharmacologically effective and suitable for contact withthe tissues of subjects without undue toxicity, irritation, or allergicresponse. A compound described herein may possess a sufficiently acidicgroup, a sufficiently basic group, both types of functional groups, ormore than one of each type, and accordingly react with a number ofinorganic or organic bases, and inorganic and organic acids, to form apharmaceutically acceptable salt. Such salts include:

(1) acid addition salts, which can be obtained by reaction of the freebase of the parent compound with inorganic acids such as hydrochloricacid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, andperchloric acid and the like, or with organic acids such as acetic acid,oxalic acid, (D) or (L) malic acid, maleic acid, methane sulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaricacid, citric acid, succinic acid or malonic acid and the like; or

(2) salts formed when an acidic proton present in the parent compoundeither is replaced by a metal ion, e.g., an alkali metal ion, analkaline earth ion, or an aluminum ion; or coordinates with an organicbase such as ethanolamine, diethanolamine, triethanolamine,trimethamine, N-methylglucamine, and the like.

Pharmaceutically acceptable salts are well known to those skilled in theart, and any such pharmaceutically acceptable salt may be contemplatedin connection with the embodiments described herein. Examples ofpharmaceutically acceptable salts include sulfates, pyrosulfates,bisulfates, sulfites, bisulfites, phosphates, monohydrogen-phosphates,dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides,bromides, iodides, acetates, propionates, decanoates, caprylates,acrylates, formates, isobutyrates, caproates, heptanoates, propiolates,oxalates, malonates, succinates, suberates, sebacates, fumarates,maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates,chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates,methoxybenzoates, phthalates, sulfonates, methylsulfonates,propylsulfonates, besylates, xylenesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, phenylacetates, phenylpropionates,phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycolates,tartrates, and mandelates. Lists of other suitable pharmaceuticallyacceptable salts are found in Remington's Pharmaceutical Sciences, 17thEdition, Mack Publishing Company, Easton, Pa., 1985.

For a compound of Formula I-IIIA that contains a basic nitrogen, apharmaceutically acceptable salt may be prepared by any suitable methodavailable in the art, for example, treatment of the free base with aninorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuricacid, sulfamic acid, nitric acid, boric acid, phosphoric acid, and thelike, or with an organic acid, such as acetic acid, phenylacetic acid,propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid,hydroxymaleic acid, isethionic acid, succinic acid, valeric acid,fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid,salicylic acid, oleic acid, palmitic acid, lauric acid, a pyranosidylacid, such as glucuronic acid or galacturonic acid, an alpha-hydroxyacid, such as mandelic acid, citric acid, or tartaric acid, an aminoacid, such as aspartic acid or glutamic acid, an aromatic acid, such asbenzoic acid, 2-acetoxybenzoic acid, naphthoic acid, or cinnamic acid, asulfonic acid, such as laurylsulfonic acid, p-toluenesulfonic acid,methanesulfonic acid, or ethanesulfonic acid, or any compatible mixtureof acids such as those given as examples herein, and any other acid andmixture thereof that are regarded as equivalents or acceptablesubstitutes in light of the ordinary level of skill in this technology.

The disclosure also relates to pharmaceutically acceptable prodrugs ofthe compounds of Formula I-IIIA, and treatment methods employing suchpharmaceutically acceptable prodrugs. The term “prodrug” means aprecursor of a designated compound that, following administration to asubject, yields the compound in vivo via a chemical or physiologicalprocess such as solvolysis or enzymatic cleavage, or under physiologicalconditions (e.g., a prodrug on being brought to physiological pH isconverted to the compound of Formula I-IIIA). A “pharmaceuticallyacceptable prodrug” is a prodrug that is non-toxic, biologicallytolerable, and otherwise biologically suitable for administration to thesubject. Illustrative procedures for the selection and preparation ofsuitable prodrug derivatives are described, for example, in “Design ofProdrugs”, ed. H. Bundgaard, Elsevier, 1985.

The present disclosure also relates to pharmaceutically activemetabolites of compounds of Formula I-IIIA, and uses of such metabolitesin the methods of the disclosure. A “pharmaceutically active metabolite”means a pharmacologically active product of metabolism in the body of acompound of Formula I-IIIA, or salt thereof. Prodrugs and activemetabolites of a compound may be determined using routine techniquesknown or available in the art. See, e.g., Bertolini et al., J. Med.Chem. 1997, 40, 2011-2016; Shan et al., J Pharm. Sci. 1997, 86 (7),765-767; Bagshawe, Drug Dev. Res. 1995, 34, 220-230; Bodor, Adv. DrugRes. 1984, 13, 255-331; Bundgaard, Design of Prodrugs (Elsevier Press,1985); and Larsen, Design and Application of Prodrugs, Drug Design andDevelopment (Krogsgaard-Larsen et al., eds., Harwood AcademicPublishers, 1991).

Any formula depicted herein is intended to represent a compound of thatstructural formula as well as certain variations or forms. For example,a formula given herein is intended to include a racemic form, or one ormore enantiomeric, diastereomeric, or geometric isomers, or a mixturethereof. Additionally, any formula given herein is intended to referalso to a hydrate, solvate, or polymorph of such a compound, or amixture thereof. For example, it will be appreciated that compoundsdepicted by a structural formula containing the symbol “

” include both stereoisomers for the carbon atom to which the symbol “

” is attached, specifically both the bonds “

” and “

” are encompassed by the meaning of “

”. For example, in some exemplary embodiments, certain compoundsprovided herein can be described by the formula

which formula will be understood to encompass compounds having bothstereochemical configurations at the relevant carbon atom, specificallyin this example

and other stereochemical combinations depending on the identity of eachR³ and R^(3′).

Any formula given herein is also intended to represent unlabeled formsas well as isotopically labeled forms of the compounds. Isotopicallylabeled compounds have structures depicted by the formulas given hereinexcept that one or more atoms are replaced by an atom having a selectedatomic mass or mass number. Examples of isotopes that can beincorporated into compounds of the disclosure include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, andiodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O ³¹P, ³²P, ³⁵S ¹⁸F,³⁶Cl, and ¹²⁵I, respectively. Such isotopically labelled compounds areuseful in metabolic studies (preferably with ¹⁴C), reaction kineticstudies (with, for example ²H or ³H), detection or imaging techniques[such as positron emission tomography (PET) or single-photon emissioncomputed tomography (SPECT)] including drug or substrate tissuedistribution assays, or in radioactive treatment of subjects. Further,substitution with heavier isotopes such as deuterium (i.e., ²H) mayafford certain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements. Isotopically labeled compounds of this disclosure andprodrugs thereof can generally be prepared by carrying out theprocedures disclosed in the schemes or in the examples and preparationsdescribed below by substituting a readily available isotopically labeledreagent for a non-isotopically labeled reagent.

As used herein, “ALK” refers to anaplastic lymphoma kinase, which alongwith leukocyte tyrosine kinase (LTK), belongs to the insulin receptor(IR) superfamily of receptor tyrosine kinases. It will be appreciatedthat ALK refers to the ALK gene, the corresponding mRNA transcribed fromthe ALK gene, the protein product of translation of the correspondingmRNA, as well as each of the aforementioned involving the rearrangementor fusion of a portion of ALK with another gene or gene product,including but not limited to NPM, EML4, TPR, TFG, ATIC, CLTC1, TPM4,MSN, ALO17, MYH9, and the like. Furthermore, it will be appreciated that“ALK” refers to mutations in the ALK gene, or the ALK protein, that canbe the result of acquired resistance mechanisms to treatment with ALKinhibitors, as described above. It will be appreciated that the ALKmutation is not necessarily dependent on the identity of therearrangement or fusion partner, e.g. EML4, NPM, and the like, and thatthe mutation can be, for example a missense mutation, an insertion, or adeletion that occurs in the ALK portion of the ALK rearrangement orfusion protein. Examples of mutation sites include, but are not limitedto L1196, L1198, G1202, D1203, S1206, T1151, L1152, E1210, F1174, C1156,I1171, V1180, F1245, G1269, R1275, and the like, and combinationsthereof. Examples of ALK mutations include but are not limited toL1196M, G1202R, C1156Y, D1203N, G1202 deletion, E1210K, S1206C, F1174C,F1174L, F1174S, F1174V, F1245C, G1269A, G1269S, I1171N, L1152P, L1152R,L1198F, R1275Q, S1206R, T1151-L1152insT, T1151M, V1180L, and the like,and combinations thereof. Examples of multiple ALK mutations include butare not limited to E1210K/D1203N, E1210K/S1206C, L1198F/C1156Y,L1198F/G1202R, L1198F/L1196M, L1196M/G1202R, L1198F/C₁₁₅₆Y,G1202R/G1269A, G1202R/G1269A/L1204V and the like.

As used herein, the term “cancer” includes, but is not limited to, ALCL,NSCLC, neuroblastoma, inflammatory myofibroblastic tumor, adult renalcell carcinoma, pediatric renal cell carcinoma, breast cancer, ER⁺breast cancer, colonic adenocarcinoma, glioblastoma, glioblastomamultiforme, anaplastic thyroid cancer, cholangiocarcinoma, ovariancancer, gastric adenocarcinoma, colorectal cancer, inflammatorymyofibroblastic tumor, angiosarcoma, epithelioid hemangioendothelioma,intrahepatic cholangiocarcinoma, thyroid papillary cancer, spitzoidneoplasms, sarcoma, astrocytoma, brain lower grade glioma, secretorybreast carcinoma, mammary analogue carcinoma, acute myeloid leukemia,congenital mesoblastic nephroma, congenital fibrosarcomas, Ph-like acutelymphoblastic leukemia, thyroid carcinoma, skin cutaneous melanoma, headand neck squamous cell carcinoma, pediatric glioma CML, prostate cancer,lung squamous carcinoma, ovarian serous cystadenocarcinoma, skincutaneous melanoma, castrate-resistant prostate cancer, Hodgkinlymphoma, and serous and clear cell endometrial cancer.

Representative Embodiments

In some embodiments, each R¹ and R² is independently H, deuterium,halogen, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3-to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclicheteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a), —OC(O)NR^(a)R^(b),—OS(O)R^(a), —OS(O)₂R^(a), —SR^(a), —S(O)R^(a), —S(O)₂R^(a),—S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b), —OS(O)NR^(a)R^(b),—OS(O)₂NR^(a)R^(b), —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b),—NR^(a)S(O)NR^(a)R^(b), —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a),—C(O)OR^(a), —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),—P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b), —P(O)OR^(a),—P(O)₂OR^(a), —CN, or —NO₂, wherein each hydrogen atom in C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, and mono- or bicyclic heteroaryl isindependently optionally substituted by deuterium, halogen, C₁-C₆ alkyl,C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),—OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),—OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f),—S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂. In some embodiments, R¹ and R² takentogether with the carbon atom to which they are attached combine to forma C₃-C₆ cycloalkyl. In some embodiments, each R¹ and R², when present,is independently H, deuterium, C₁-C₆ alkyl.

In some embodiments, each R³ and R^(3′) is independently H, deuterium,halogen, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3-to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclicheteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a), —OC(O)NR^(a)R^(b),—OS(O)R^(a), —OS(O)₂R^(a), —SR^(a), —S(O)R^(a), —S(O)₂R^(a),—S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b), —OS(O)NR^(a)R^(b),—OS(O)₂NR^(a)R^(b), —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b),—NR^(a)S(O)NR^(a)R^(b), —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a),—C(O)OR^(a), —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),—P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b), —P(O)OR^(a),—P(O)₂OR^(a), —CN, or —NO₂, wherein each hydrogen atom in C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic heteroaryl, isindependently optionally substituted by deuterium, halogen, C₁-C₆ alkyl,C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),—OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),—OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f),—S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂. In some embodiments, R³ and R^(3′) takentogether with the carbon atom to which they are attached combine to forma C₃-C₆ cycloalkyl, wherein each hydrogen atom in C₃-C₆ cycloalkyl isindependently optionally substituted by deuterium, halogen, C₁-C₆ alkyl,C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),—OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),—OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f),—S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂. In some embodiments, each R³ and R^(3′) isindependently H, deuterium, C₁-C₆ alkyl. In some embodiments, R³ andR^(3′) taken together with the carbon atom to which they are attachedcombine to form a C₃-C₆ cycloalkyl. In some embodiments, each R^(3′) isH or deuterium. In some embodiments, both R^(3′) taken together combineto form a divalent group —(CR¹R²)_(m)—, where m is 1 or 2.

In some embodiments, each R⁴ and R⁵ is independently hydrogen,deuterium, halogen, —OR^(c), —OC(O)R^(c), —OC(O)NR^(c)R^(d),—OC(═N)NR^(c)R^(d), —OS(O)R^(c), —OS(O)₂R^(c), —OS(O)NR^(c)R^(d),—OS(O)₂NR^(c)R^(d), —SR^(c), —S(O)R^(c), —S(O)₂R^(c), —S(O)NR^(c)R^(d),—S(O)₂NR^(c)R^(d), —NR^(c)R^(d), —NR^(c)C(O)R^(d), —NR^(c)C(O)OR^(d),—NR^(c)C(O)NR^(c)R^(d), —NR^(c)C(═N)NR^(c)R^(d), —NR^(c)S(O)R^(d),—NR^(c)S(O)₂R^(d), —NR^(c)S(O)NR^(c)R^(d), —NR^(c)S(O)₂NR^(c)R^(d),—C(O)R^(c), —C(O)OR^(c), —C(O)NR^(c)R^(d), —C(═N)NR^(c)R^(d),—PR^(c)R^(d), —P(O)R^(c)R^(d), —P(O)₂R^(c)R^(d), —P(O)NR^(c)R^(d),—P(O)₂NR^(c)R^(d), —P(O)OR^(c), —P(O)₂OR^(c), —CN, —NO₂, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic heteroaryl, whereineach hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- orbicyclic heteroaryl, C₅-C₈ cycloalkyl, or 5- to 8-memberedheterocycloalkyl is independently optionally substituted by deuterium,halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),—OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e),—OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e),—S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f),—NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f),—NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f),—C(O)R^(e), —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f),—P(O)R^(e)R^(f), —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),—P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂. In some embodiments, each R⁴ isH or deuterium. In some embodiments, R⁵ is fluoro.

In some embodiments, R⁶ is H, deuterium, or C₁-C₆ alkyl, wherein eachhydrogen atom in C₁-C₆ alkyl is independently optionally substituted bydeuterium, halogen, —OR^(e), —SR^(e), or —NR^(e)R^(f). In someembodiments, R⁶ is H.

In some embodiments, each R⁷ is independently hydrogen or deuterium. Insome embodiments, R⁷ is H.

In some embodiments, each R^(a), R^(b), R^(c), R^(d), R^(e), and R isindependently selected from the group consisting of H, deuterium, C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, and 5- to 7-membered heteroaryl.

In some embodiments, n is 0, 1, 2, or 3. In some embodiments, n is 0or 1. In some embodiments, n is 0. In some embodiments, n is 1.

The following represent illustrative embodiments of compounds of theFormula I-IIIA:

Cpd Structure Name 1

(4S)-4-(Difluoromethyl)-8-fluoro-13,13-dimethyl-3,4,13,14-tetrahydro-6H-18,1- (metheno)[1,4]oxazino[3,4-i]pyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-15(12H)-one a.k.a.(S,14aE,15aE)-13-(difluoromethyl)-35-fluoro-6,6-dimethyl-13,14-dihydro-12H-4-oxa-7-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclooctaphan-8-one 2

(16′S)-16′-(Difluoromethyl)-12′-fluoro-4′H,5′H,6′H,8′H,14′H,16′H,17′H-spiro[cyclopropane-1,7′-[1,19](metheno)[1,4]oxazino[3,4-j]pyrazolo[4,3-g][1,5,9,11]benzoxatriazacyclotetradecin]-4′-one a.k.a(S,4a′E,5a′E)-3′-(difluoromethyl)-5′-fluorospiro[cyclopropane-1,6′-4-oxa-8-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclononaphan]-9′-one 3

(16S)-16-(Difluoromethyl)-12-fluoro-5,6,7,8,16,17-hexahydro-4H,14H-1,19-(metheno)[1,4]oxazino[3,4- j]pyrazolo[4,3-g][1,5,9,11]benzoxatriazacyclotetradecin-4-one a.k.a(S,14aE,15aE)-13-(difluoromethyl)-35-fluoro-13,14-dihydro-12H-4-oxa-8-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclononaphan-9-one 4

(4′S)-4′-(Difluoromethyl)-8′-fluoro-3′H,4′H,6′H,12′H,14′H,15′H-spiro[cyclopropane-1,13′-[18,1](metheno)[1,4]oxazino[3,4-i]pyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecin]-15′-one a.k.a(S,4a′E,5a′E)-3′-(difluoromethyl)-5′-fluorospiro[cyclopropane-1,6′-4-oxa-7-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclooctaphan]-8′-one 5

(6R,16S)-16-(Difluoromethyl)-12-fluoro-6-methyl-5,6,7,8,16,17-hexahydro-4H,14H-1,19-(metheno)[1,4]oxazino[3,4-j]pyrazolo[4,3-g][1,5,9,11]benzoxatriazacyclotetradecin-4-one a.k.a(13S,14aE,15aE,7R)-13-(difluoromethyl)-35-fluoro-7-methyl-13,14-dihydro-12H-4-oxa-8-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclononaphan-9-one 6

(16′S)-16′-(Difluoromethyl)-12′-fluoro-4′H,5′H,7′H,8′H,14′H,16′H,17′H-spiro[cyclopropane-1,6′-[1,19](metheno)[1,4]oxazino[3,4-j]pyrazolo[4,3-g][1,5,9,11]benzoxatriazacyclotetradecin]-4′-one a.k.a.(S,4a′E,5a′E)-3′-(difluoromethyl)-5′-fluorospiro[cyclopropane-1,7′-4-oxa-8-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclononaphan]-9′-one 7

(16S)-16-(Difluoromethyl)-12-fluoro-7,7-dimethyl-5,6,7,8,16,17-hexahydro-4H,14H-1,19-(metheno)[1,4]oxazino[3,4-j]pyrazolo[4,3-g][1,5,9,11]benzoxatriazacyclotetradecin-4-one a.k.a.(S,14aE,15aE)-13-(difluoromethyl)-35-fluoro-6,6-dimethyl-13,14-dihydro-12H-4-oxa-8-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclononaphan-9-one 8

(16S)-16-(Difluoromethyl)-12-fluoro-6,6-dimethyl-5,6,7,8,16,17-hexahydro-4H,14H-1,19-(metheno)[1,4]oxazino[3,4-j]pyrazolo[4,3-g][1,5,9,11]benzoxatriazacyclotetradecin-4-one a.k.a.(S,14aE,15aE)-13-(difluoromethyl)-35-fluoro-7,7-dimethyl-13,14-dihydro-12H-4-oxa-8-aza-1(4,6)-pyrazolo[1′,5′:1,2]pyrimido[5,4-b][1,4]oxazina-3(1,2)-benzenacyclononaphan-9-one

Those skilled in the art will recognize that the species listed orillustrated herein are not exhaustive, and that additional specieswithin the scope of these defined terms may also be selected.

Pharmaceutical Compositions

For treatment purposes, pharmaceutical compositions comprising thecompounds described herein may further comprise one or morepharmaceutically-acceptable excipients. A pharmaceutically-acceptableexcipient is a substance that is non-toxic and otherwise biologicallysuitable for administration to a subject. Such excipients facilitateadministration of the compounds described herein and are compatible withthe active ingredient. Examples of pharmaceutically-acceptableexcipients include stabilizers, lubricants, surfactants, diluents,anti-oxidants, binders, coloring agents, bulking agents, emulsifiers, ortaste-modifying agents. In preferred embodiments, pharmaceuticalcompositions according to the invention are sterile compositions.Pharmaceutical compositions may be prepared using compounding techniquesknown or that become available to those skilled in the art.

Sterile compositions are also contemplated by the invention, includingcompositions that are in accord with national and local regulationsgoverning such compositions.

The pharmaceutical compositions and compounds described herein may beformulated as solutions, emulsions, suspensions, or dispersions insuitable pharmaceutical solvents or carriers, or as pills, tablets,lozenges, suppositories, sachets, dragees, granules, powders, powdersfor reconstitution, or capsules along with solid carriers according toconventional methods known in the art for preparation of various dosageforms. Pharmaceutical compositions of the invention may be administeredby a suitable route of delivery, such as oral, parenteral, rectal,nasal, topical, or ocular routes, or by inhalation. Preferably, thecompositions are formulated for intravenous or oral administration.

For oral administration, the compounds the invention may be provided ina solid form, such as a tablet or capsule, or as a solution, emulsion,or suspension. To prepare the oral compositions, the compounds of theinvention may be formulated to yield a dosage of, e.g., from about 0.1mg to 1 g daily, or about 1 mg to 50 mg daily, or about 50 to 250 mgdaily, or about 250 mg to 1 g daily. Oral tablets may include the activeingredient(s) mixed with compatible pharmaceutically acceptableexcipients such as diluents, disintegrating agents, binding agents,lubricating agents, sweetening agents, flavoring agents, coloring agentsand preservative agents. Suitable inert fillers include sodium andcalcium carbonate, sodium and calcium phosphate, lactose, starch, sugar,glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol, andthe like. Exemplary liquid oral excipients include ethanol, glycerol,water, and the like. Starch, polyvinyl-pyrrolidone (PVP), sodium starchglycolate, microcrystalline cellulose, and alginic acid are exemplarydisintegrating agents. Binding agents may include starch and gelatin.The lubricating agent, if present, may be magnesium stearate, stearicacid, or talc. If desired, the tablets may be coated with a materialsuch as glyceryl monostearate or glyceryl distearate to delay absorptionin the gastrointestinal tract, or may be coated with an enteric coating.

Capsules for oral administration include hard and soft gelatin capsules.To prepare hard gelatin capsules, active ingredient(s) may be mixed witha solid, semi-solid, or liquid diluent. Soft gelatin capsules may beprepared by mixing the active ingredient with water, an oil, such aspeanut oil or olive oil, liquid paraffin, a mixture of mono anddi-glycerides of short chain fatty acids, polyethylene glycol 400, orpropylene glycol.

Liquids for oral administration may be in the form of suspensions,solutions, emulsions, or syrups, or may be lyophilized or presented as adry product for reconstitution with water or other suitable vehiclebefore use. Such liquid compositions may optionally contain:pharmaceutically-acceptable excipients such as suspending agents (forexample, sorbitol, methyl cellulose, sodium alginate, gelatin,hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel andthe like); non-aqueous vehicles, e.g., oil (for example, almond oil orfractionated coconut oil), propylene glycol, ethyl alcohol, or water;preservatives (for example, methyl or propyl p-hydroxybenzoate or sorbicacid); wetting agents such as lecithin; and, if desired, flavoring orcoloring agents.

For parenteral use, including intravenous, intramuscular,intraperitoneal, intranasal, or subcutaneous routes, the agents of theinvention may be provided in sterile aqueous solutions or suspensions,buffered to an appropriate pH and isotonicity or in parenterallyacceptable oil. Suitable aqueous vehicles include Ringer's solution andisotonic sodium chloride. Such forms may be presented in unit-dose formsuch as ampoules or disposable injection devices, in multi-dose formssuch as vials from which the appropriate dose may be withdrawn, or in asolid form or pre-concentrate that can be used to prepare an injectableformulation. Illustrative infusion doses range from about 1 to 1000μg/kg/minute of agent admixed with a pharmaceutical carrier over aperiod ranging from several minutes to several days.

For nasal, inhaled, or oral administration, the inventive pharmaceuticalcompositions may be administered using, for example, a spray formulationalso containing a suitable carrier. The inventive compositions may beformulated for rectal administration as a suppository.

For topical applications, the compounds of the present invention arepreferably formulated as creams or ointments or a similar vehiclesuitable for topical administration. For topical administration, theinventive compounds may be mixed with a pharmaceutical carrier at aconcentration of about 0.1% to about 10% of drug to vehicle. Anothermode of administering the agents of the invention may utilize a patchformulation to effect transdermal delivery.

As used herein, the terms “treat” or “treatment” encompass both“preventative” and “curative” treatment. “Preventative” treatment ismeant to indicate a postponement of development of a disease, a symptomof a disease, or medical condition, suppressing symptoms that mayappear, or reducing the risk of developing or recurrence of a disease orsymptom. “Curative” treatment includes reducing the severity of orsuppressing the worsening of an existing disease, symptom, or condition.Thus, treatment includes ameliorating or preventing the worsening ofexisting disease symptoms, preventing additional symptoms fromoccurring, ameliorating or preventing the underlying systemic causes ofsymptoms, inhibiting the disorder or disease, e.g., arresting thedevelopment of the disorder or disease, relieving the disorder ordisease, causing regression of the disorder or disease, relieving acondition caused by the disease or disorder, or stopping the symptoms ofthe disease or disorder.

The term “subject” refers to a mammalian patient in need of suchtreatment, such as a human, or, in the case of veterinary applications,can be a laboratory, agricultural, or domestic animal. In someembodiments, the subject can be a human. In some embodiments, thesubject can be a domestic animal for veterinary uses, such as a dog orcat. In some embodiments, the subject can be an agricultural animal,such as a cow, pig, horse, and the like. In some embodiments, thesubject can be a laboratory animal such as a rodent (e.g. mouse, rat,etc), primate (e.g. rhesus monkey, and the like), a canine, and thelike.

Exemplary diseases include cancer, pain, neurological diseases,autoimmune diseases, and inflammation. Cancer includes, for example,lung cancer, colon cancer, breast cancer, prostate cancer,hepatocellular carcinoma, renal cell carcinoma, gastric andesophago-gastric cancers, glioblastoma, head and neck cancers,inflammatory myofibroblastic tumors, and anaplastic large cell lymphoma.Pain includes, for example, pain from any source or etiology, includingcancer pain, pain from chemotherapeutic treatment, nerve pain, pain frominjury, or other sources. Autoimmune diseases include, for example,rheumatoid arthritis, Sjogren syndrome, Type I diabetes, and lupus.Exemplary neurological diseases include Alzheimer's Disease, Parkinson'sDisease, Amyotrophic lateral sclerosis, and Huntington's disease.Exemplary inflammatory diseases include atherosclerosis, allergy, andinflammation from infection or injury.

In one aspect, the compounds and pharmaceutical compositions of theinvention specifically target tyrosine receptor kinases, in particularALK, more particularly ALK having one or more mutations as describedherein. Thus, these compounds and pharmaceutical compositions can beused to prevent, reverse, slow, or inhibit the activity of ALK. Inpreferred embodiments, methods of treatment target cancer. In otherembodiments, methods are for treating lung cancer, particularlynon-small cell lung cancer.

In the inhibitory methods of the invention, an “effective amount” meansan amount sufficient to inhibit the target protein. Measuring suchtarget modulation may be performed by routine analytical methods such asthose described below. Such modulation is useful in a variety ofsettings, including in vitro assays. In such methods, the cell ispreferably a cancer cell with abnormal signaling due to upregulation ofALK, in particular particularly ALK having one or more mutations asdescribed herein.

In some embodiments, the uses and methods described herein for anycompound for the Formula I-IIIa can be for treating cancer mediated byALK or mediated by a rearrangement or fusion of ALK with another gene orgene product, including but not limited to NPM, EML4, TPR, TFG, ATIC,CLTC1, TPM4, MSN, ALO17, MYH9, and the like. In some embodiments, theuses and methods described herein for any compound for the FormulaI-IIIa can be for treating cancer mediated by ALK or mediated by arearrangement or fusion of ALK with another gene or gene product,including but not limited to NPM, EML4, TPR, TFG, ATIC, CLTC1, TPM4,MSN, ALO17, MYH9, and the like, and having at least one mutation, suchas a missense mutation, an insertion, or a deletion that occurs in theALK portion of the ALK rearrangement or fusion protein, wherein themutation site includes, but is not limited to L1196, L1198, G1202,D1203, S1206, T1151, L1152, E1210, F1174, C1156, I1171, V1180, F1245,G1269, R1275, and the like, and combinations thereof.

In some embodiments, the uses and methods described herein for anycompound for the Formula I-IIIa can be for treating cancer mediated byALK or mediated by a rearrangement or fusion of ALK with another gene orgene product, including but not limited to NPM, EML4, TPR, TFG, ATIC,CLTC1, TPM4, MSN, ALO17, MYH9, and the like, and having at least onemutation, such as a missense mutation, an insertion, or a deletion thatoccurs in the ALK portion of the ALK rearrangement or fusion protein,wherein the ALK mutation is L1196M, G1202R, C1156Y, D1203N, G1202deletion, E1210K, S1206C, F1174C, F1174L, F1174S, F1174V, F1245C,G1269A, G1269S, I1171N, L1152P, L1152R, L1198F, R1275Q, S1206R,T1151-L1152insT, T1151M, V1180L, or the like, and combinations thereof.In some embodiments, the uses and methods described herein for anycompound for the Formula I-IIIa can be for treating cancer mediated byALK or mediated by a rearrangement or fusion of ALK with another gene orgene product, including but not limited to NPM, EML4, TPR, TFG, ATIC,CLTC1, TPM4, MSN, ALO17, MYH9, and the like, and having at least onemutation, such as a missense mutation, an insertion, or a deletion thatoccurs in the ALK portion of the ALK rearrangement or fusion protein,wherein the mutation is a multiple ALK mutation selected fromE1210K/D1203N, E1210K/S1206C, L1198F/C1156Y, L1198F/G1202R,L1198F/L1196M, L1196M/G1202R, L1198F/C1156Y, G1202R/G1269A, andG1202R/G1269A/L1204V.

In treatment methods according to the invention, an “effective amount”means an amount or dose sufficient to generally bring about the desiredtherapeutic benefit in subjects needing such treatment. Effectiveamounts or doses of the compounds of the invention may be ascertained byroutine methods, such as modeling, dose escalation, or clinical trials,taking into account routine factors, e.g., the mode or route ofadministration or drug delivery, the pharmacokinetics of the agent, theseverity and course of the infection, the subject's health status,condition, and weight, and the judgment of the treating physician. Anexemplary dose is in the range of about from about 0.1 mg to 1 g daily,or about 1 mg to 50 mg daily, or about 50 to 250 mg daily, or about 250mg to 1 g daily. The total dosage may be given in single or divideddosage units (e.g., BID, TID, QID).

Once improvement of the subject's disease has occurred, the dose may beadjusted for preventative or maintenance treatment. For example, thedosage or the frequency of administration, or both, may be reduced as afunction of the symptoms, to a level at which the desired therapeutic orprophylactic effect is maintained. Of course, if symptoms have beenalleviated to an appropriate level, treatment may cease. Subjects may,however, require intermittent treatment on a long-term basis upon anyrecurrence of symptoms. Subjects may also require chronic treatment on along-term basis.

Drug Combinations

The inventive compounds described herein may be used in pharmaceuticalcompositions or methods in combination with one or more additionalactive ingredients in the treatment of the diseases and disordersdescribed herein. Further additional active ingredients include othertherapeutics or agents that mitigate adverse effects of therapies forthe intended disease targets. Such combinations may serve to increaseefficacy, ameliorate other disease symptoms, decrease one or more sideeffects, or decrease the required dose of an inventive compound. Theadditional active ingredients may be administered in a separatepharmaceutical composition from a compound of the present invention ormay be included with a compound of the present invention in a singlepharmaceutical composition. The additional active ingredients may beadministered simultaneously with, prior to, or after administration of acompound of the present invention.

Combination agents include additional active ingredients are those thatare known or discovered to be effective in treating the diseases anddisorders described herein, including those active against anothertarget associated with the disease. For example, compositions andformulations of the invention, as well as methods of treatment, canfurther comprise other drugs or pharmaceuticals, e.g., other activeagents useful for treating or palliative for the target diseases orrelated symptoms or conditions. For cancer indications, additional suchagents include, but are not limited to, kinase inhibitors, such as EGFRinhibitors (e.g., erlotinib, gefitinib), Raf inhibitors (e.g.,vemurafenib), VEGFR inhibitors (e.g., sunitinib), ALK inhibitors (e.g.,crizotinib) standard chemotherapy agents such as alkylating agents,antimetabolites, anti-tumor antibiotics, topoisomerase inhibitors,platinum drugs, mitotic inhibitors, antibodies, hormone therapies, orcorticosteroids. For pain indications, suitable combination agentsinclude anti-inflammatories such as NSAIDs. The pharmaceuticalcompositions of the invention may additionally comprise one or more ofsuch active agents, and methods of treatment may additionally compriseadministering an effective amount of one or more of such active agents.

Chemical Synthesis

Exemplary chemical entities useful in methods of the description willnow be described by reference to illustrative synthetic schemes fortheir general preparation below and the specific examples that follow.Artisans will recognize that, to obtain the various compounds herein,starting materials may be suitably selected so that the ultimatelydesired substituents will be carried through the reaction scheme with orwithout protection as appropriate to yield the desired product.Alternatively, it may be necessary or desirable to employ, in the placeof the ultimately desired substituent, a suitable group that may becarried through the reaction scheme and replaced as appropriate with thedesired substituent. Furthermore, one of skill in the art will recognizethat the transformations shown in the schemes below may be performed inany order that is compatible with the functionality of the particularpendant groups.

Abbreviations

The examples described herein use materials, including but not limitedto, those described by the following abbreviations known to thoseskilled in the art:

g grams eq equivalents mmol millimoles mL milliliters EtOAc or EA ethylacetate MHz megahertz Ppm parts per million Δ chemical shift S singlet Ddoublet T triplet Q quartet Quin quintet Br broad M multiplet Hz hertzTHF tetrahydrofuran ° C. degrees Celsius R_(f) retardation factor Nnormal J coupling constant DMSO-d₆ deuterated dimethyl sulfoxide EtOHethanol DIPEA N,N-diisopropylethylamine min minutes hr hours TLC thinlayer chromatography M molar MS mass spectrum m/z mass-to-charge ratioDMAP 4-(dimethylamino)pyridine μM micromolar IC₅₀ half maximalinhibitory concentration U/mL units of activity per milliliter MOM-Clmethoxymethyl chloride DCM dichloromethane DMF N,N-methylformamide

EXAMPLES Comparative Examples

Comparative Examples 1 and 2 were made as described in International PCTPublication No. WO 2019126122, corresponding to International PCTApplication No. PCT/US2018/066159, filed Dec. 18, 2018.

Comparative Example Structure Comp. Ex. 1

Comp. Ex. 2

Example 1: General Method A Preparation of tert-Butyl{1-[2-(chloromethyl)-4-fluorophenoxy]-2-methylpropan-2-yl}carbamate(A-1)

Step 1: To A-1-1 (259 mg, 1.52 mmol) in anhydrous DMF (7.35 mL) wasadded NaH (60% in mineral oil, 91.3 mg, 2.28 mmol) under argon at 0° C.The reaction was allowed to stir and warm over 1 hr. The reaction wascooled to 0° C. and A-1-2 was added. The reaction was stirred and thetemperature was increased to 50° C. and stirred for 18 hr. The reactionwas quenched with saturated NH₄Cl (aq. 5 mL) at 0° C. and stirredvigorously. Water (25 mL) was added and extracted with DCM (3×25 mL).The combined organic layer was washed with brine and dried over sodiumsulfate. Flash column chromatography (12 g silica, ISCO, 0-50% EA inHexanes) provided A-1-3 (267 mg, 51% yield).

Step 2: to A-1-3 (267 mg, 0.782 mmol) in anhydrous THF (3.9 mL) wasadded LiBH₄ (34 mg, 1.56 mmol) at 0° C. The reaction was stirred for 18hr as temperature increased to ambient. The reaction was quenched withwater (5 mL) and 2 M NaOH (1 mL), then extracted with DCM (3×10 mL). Thecombined organic layer was washed with brine and dried over sodiumsulfate. Flash column chromatography (12 g silica, ISCO, 0-50% EA inHexanes with ELSD detection) provided A-1-4 (29.3 mg, 12% yield).

Step 3: To A-1-4 (29.3 mg, 93.5 μmol) in DCM (1 mL) was added Hunig'sbase (36.2 mg, 280.5 μmol, 48.8 μL). The reaction was cooled to 0° C.and mesyl chloride (13.9 mg, 121.5 μmol, 9.4 uL) was added. The reactionwas stirred as temperature increase to 0-22° C. over 18 hr. The reactionwas quenched with 2 M HCl (aq) (1 mL) at 0° C. Diluted with water andDCM (5 mL each), layers were partitioned and the aqueous layer wasextracted DCM (2×5 mL). The combined organic layer was washed with brineand dried over sodium sulfate. Flash column chromatography (12 g silica,ISCO, 0-50% EA in Hex) to afford A-1 (25.2 mg, 75.9 μmol, 81% yield).

Example 2: General Method B Preparation of tert-Butyl[(1-{[2-(chloromethyl)-4-fluorophenoxy]methyl}cyclopropyl)methyl]carbamate (A-2)

Step 1: To a solution of A-1-1 (100 mg, 587.7 μmol), A-2-1 (118.2 mg,587.7 μmol), and PPh₃ (231.2 mg, 881.6 μmol) in anhydrous DCM (403.14μL) at 0° C. was added DIAD (190.2 mg, 940.4 μmol, 184.6 μL) withstirring. The mixture was stirred for 18 hr as it warmed to RT. Flashcolumn chromatography (ISCO, 12 g silica, 0-50% ethyl acetate inhexanes) provided A-2-2 (163.1 mg, 433.3 μmol, 73.7% yield)

Step 2: Compound A-2 was prepared according to General Method A startingwith B-1-2 in step 2.

Compound A-3 was prepared according to General Method B

Compound A-5, A-6, A-7, and A-8 were prepared according to GeneralMethod B

Example 3: General Method C Preparation of tert-butyl(1-((2-(chloromethyl)-4-fluorophenoxy)methyl)cyclopropyl)carbamate (A-4)

Compound A-4 was prepared according to General Method C

Step 1. To a solution of A-4-1 (500 mg, 2.67 mmol) in DCM (12.25 mL) wasadded triethylamine (811 mg, 8.0 mmol, 1.12 mL). The mixture was cooledto 0° C. and p-toluenesulfonyl chloride (TosCl, 611 mg, 3.2 mmol) andDMAP (6.5 mg, 53 μmol) were added. The reaction was warmed to 22° C. andstirred for 18 hr. Water (15 mL) and DCM (10 mL) were added and theorganic layer separated. The aqueous layer was extracted with DCM (2×10mL), dried with Na₂SO₄ and concentrated under reduced pressure. Flashchromatography (ISCO system, silica 24 g, 0-30% ethyl acetate in hexane)provided A-4-2 (608 mg, 1.78 mmol, 67%).

Step 2. To a solution of A-1-1 (100 mg, 588 μmol) in DMF (3.00 L) wasadded cesium carbonate (479 mg, 1.5 mmol) and A-4-2 (200 mg, 588 mol).The mixture was stirred at 70° C. for 17 hr. The reaction mixture wascooled, diluted with DCM (5 mL), filtered through a syringe filter andconcentrated under reduced pressure. Flash column chromatography (12 gsilica, ISCO, 0-80% ethyl acetate in hexane) gave A-4-3 (47 mg, 138μmol, 23% yield).

Step 3. To A-4-3 (47 mg, 138 μmol) in anhydrous THF (3.9 mL) at 0° C.was added LiBH₄ (9 mg, 415 μmol). The reaction was warmed to 22° C. andstirred for 21 hrs. The reaction was cooled to 0° C. and quenched withand 2 M NaOH (3 mL) and water (3 mL) then extracted with DCM (3×5 mL).The combined organic layer was washed with brine and dried over sodiumsulfate and concentrated under reduced pressure. Flash columnchromatography (12 g silica, ISCO, 0-12.5% methanol in dichloromethane)provided A-4-4 (37.1 mg, 119 μmol, 86% yield).

Step 4. To A-4-4 (37.1 mg, 120 μmol) in DCM (1 mL) was added mesylchloride (20.5 mg, 179 μmol, 14 uL). The reaction was cooled to 0° C.and Hunig's base (46 mg, 357 μmol, 62 μL) was added. The reaction wasslowly warmed to 22° C. and stirred for 71 hr. The reaction was cooledto −20° C. and quenched with 2 M HCl (aq) (2 mL). Diluted with water andDCM (2 mL each), layers were partitioned, and the aqueous layer wasextracted DCM (2×5 mL). The combined organic layer was washed with brineand dried over sodium sulfate. Flash column chromatography (12 g silica,ISCO, 0-40% ethyl acetate in hexane) to afford A-4 (25.2 mg, 75.9 μmol,75% yield).

MS [M + Na] Cpd # Structure (R = OMs, Cl) m/z A-1

354.1 A-2

366.1 A-3

340.1 A-4

352.1 A-5

354.1 A-6

366.1 A-7

368.2 A-8

Example 4: General Method D Preparation of ethyl5-chloro-6-hydroxy-pyrazolo[1,5-a]pyrimidine-3-carboxylate (B)

Step 1. To a solution of B-1 (150 g, 1.44 mol, 143 mL, 1.00 eq.) andB-1A (104 g, 1.73 mol, 105 mL, 1.20 eq.) in tetrahydrofuran (3.00 L) wasadded sodium hydride (80.7 g, 2.02 mol, 60.0% purity, 1.40 eq.) slowlyat 0° C. over a period of 30 minutes under nitrogen. During which thetemperature was maintained below 0° C. The reaction mixture was stirredat 0° C. for 12 hr. The formation of white solids was observed, methyltert-butyl ether (2.00 L) was added, filtered, and the filtered cake wasdried under reduced pressure to give the crude B-2 (283 g, crude) aslight-yellow solid.

Step 2. To a solution of B-2A (165 g, 1.06 mol, 1.00 eq.) in DMF (3.00L) was added cesium carbonate (624 g, 1.91 mol, 1.80 eq.) and B-2 (279g, 1.91 mol, 1.80 eq.). The mixture was stirred at 110° C. for 12 hr.The reaction mixture was diluted with water (3.00 L), hydrochloric acid(5.00 M, 1.80 L) was added to the mixture slowly at 20° C., and theresulting precipitated solids was filtered and washed with methylalcohol (300 mL). The filtered cake was concentrated under reducedpressure to give the crude B-3 (162 g, 609 mmol, 57.4% yield, 89.3%purity) as yellow solid.

Step 3. B-3 (100 g, 375 mmol, 1.00 eq.) was added into phosphorusoxychloride (300 mL). The mixture was stirred at 110° C. for 12 hr. Thereaction mixture was concentrated under reduced pressure to removesolvent and until product precipitated. The residue was diluted with icewater (1.00 L) and filtered to remove the solvent. Then the filter cakewas dissolved in dichloromethane (2.00 L) and water (2.00 L) was added.The organic phase was separated, washed with brine (300 mL), dried overanhydrous sodium sulfate, filtered and concentrated under reducedpressure to give B-4 (64.0 g, 213 mmol, 56.7% yield, 85.0% purity) asgray solid.

Step 4. Aluminum trichloride (752 g, 5.64 mol, 308 mL, 5.00 eq.) wasadded in one portion to anhydrous dichloroethane (4.90 L) and themixture was stirred under nitrogen at 20° C. for 10 minutes, then B-4(324 g, 1.13 mol, 1.00 eq.) was added to the mixture in five equalportions. The mixture was stirred at 20° C. for 24 hr. The reactionmixture was quenched by addition of hydrochloric acid (5.00 M, 2.00 L)at 0° C., diluted with water (1.00 L), and then extracted with ethylacetate (3.00 L×3). The combined organic layers were washed with water(2.00 L) and brine (1.00 L×2), dried over anhydrous sodium sulfate,filtered and concentrated under reduced pressure to give the crudeproduct B (280 g, 920 mmol, 81.6% yield, 79.4% purity) as gray solid.

Example 5: General Method E Preparation of Compound ethyl(3S)-3-(difluoromethyl)-3,4-dihydro-2H-pyrazolo[1,2]pyrimido[2,4-d][1,4]oxazine-6-carboxylate(C)

Step 1: To a solution of C-1 (5.00 g, 22.1 mmol, 1.00 eq.) in DCM (120mL) was added pyridine (2.80 g, 35.4 mmol, 2.85 mL, 1.60 eq.) andtrifluoroacetic anhydride (7.48 g, 26.5 mmol, 4.37 mL, 1.20 eq.) at −20°C. The mixture was stirred at −20° C.-0° C. for 5 hr. The reactionmixture was concentrated under reduced pressure to remove solvent at 30°C. The residue was diluted with water (50.0 mL) and extracted withPetroleum ether/Ethyl acetate=10:1 (50.0 mL×3). The combined organiclayers were washed with brine (30.0 mL), dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (SiO₂,Petroleum ether/Ethyl acetate=1/0 to 50/1) to give C-2 (6.70 g, 16.8mmol, 76.2% yield, 90.0% purity) as a light-yellow oil.

Step 2. To a solution of C-2 (0.50 g, 1.40 mmol, 1.00 eq.) in dioxane(5.00 mL) was added (4-methoxyphenyl)methanamine (229 mg, 1.67 mmol, 216μL, 1.20 eq.) and triethylamine (169 mg, 1.67 mmol, 233 μL, 1.20 eq.).The mixture was stirred at 90° C. for 12 hrs. The reaction mixture wasdiluted with water (30.0 mL) and extracted with ethyl acetate (20.0mL×2). The combined organic layers were washed with brine (15.0 mL),dried over anhydrous sodium sulfate, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, Petroleum ether/Ethyl acetate=50/1 to 10/1) togive C-3 (0.35 g, 1.01 mmol, 72.3% yield, 99.5% purity) as alight-yellow oil.

Step 3. To a solution of C-3 (3.35 g, 9.70 mmol, 1.00 eq.) intetrahydrofuran (34.0 mL) was added a solution of tetrabutyl ammoniumfluoride in tetrahydrofuran (1.00 M, 9.70 mL, 1.00 eq.). The mixture wasstirred at 20° C. for 2 hr. The reaction mixture was quenched by water(80.0 mL) at 20° C. and extracted with ethyl acetate (50.0 mL×2). Thecombined organic layers were washed with brine (50.0 mL), dried overanhydrous sodium sulfate, filtered and concentrated under reducedpressure to give a residue. The residue was purified by columnchromatography (SiO₂, Petroleum ether/Ethyl acetate=30/1 to 10/1) togive C-4 (2.10 g, 8.99 mmol, 92.7% yield, 99.0% purity) as alight-yellow oil.

Step 4. To a solution of C-4 (2.60 g, 11.2 mmol, 1.00 eq.) in dimethylsulfoxide (50.0 mL) was added potassium fluoride (1.63 g, 28.1 mmol, 658μL, 2.50 eq.) and B (2.74 g, 11.2 mmol, 1.00 eq.). The mixture wasstirred at 120° C. for 3 hr. The reaction mixture was diluted with water(300 mL) and extracted with ethyl acetate (50.0 mL×3). The combinedorganic layers were washed with brine (50.0 mL×3), dried over anhydroussodium sulfate, filtered and concentrated under reduced pressure to givea residue. The residue was purified by column chromatography (SiO₂,Petroleum ether/Ethyl acetate=10/1 to 2/1) to give compound C-5 (1.00 g,1.83 mmol, 16.3% yield, 80.3% purity) as a yellow oil.

Step 5. To a solution of C-5 (1.00 g, 2.28 mmol, 1.00 eq.) in dimethylsulfoxide (70.0 mL) was added cesium carbonate (2.97 g, 9.12 mmol, 4.00eq.). The mixture was stirred at 25° C. for 2 hr. The reaction mixturewas diluted with water (300 mL) and extracted with ethyl acetate (50.0mL×3). The combined organic layers were washed with brine (100 mL×3),dried over anhydrous sodium sulfate, filtered and concentrated underreduced pressure to give C-6 (0.75 g, 1.43 mmol, 62.9% yield, 80.0%purity) as a yellow solid.

Step 6. C-6 (0.51 g, 1.22 mmol, 1.00 eq.) was added into trifluoroaceticacid (10.0 mL). The mixture was stirred at 70° C. for 4 hr. The reactionmixture was concentrated under reduced pressure to give a residue. Theresidue was purified by column chromatography (SiO₂, Petroleumether/Ethyl acetate=10/1 to 1/1) to give C (0.27 g, 862 μmol, 70.8%yield, 96.0% purity) as a gray solid.

Example 6: General Method F Preparation of(4S)-4-(Difluoromethyl)-8-fluoro-13,13-dimethyl-3,4,13,14-tetrahydro-6H-18,1-(metheno)[1,4]oxazino[3,4-i]pyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-15(12H)-one(1)

Step 1: A-1 (R═Cl, 25.2 mg, 75.95 μmol) was dissolved in DMF (0.5 mL) atroom temperature. Cs₂CO₃ (65.55 mg, 201.18 μmol) was added followed by C(20 mg, 67.06 μmol). The mixture stirred at 22° C. for 18 hr. Reactionwas diluted with DCM (5 mL) and cooled. The solution was filtered, andthe filtrate was concentrated under reduced pressure. Flash columnchromatography (ISCO, 12 g, 20-80% ethyl acetate in hexanes) to afford1-1 (23.4 mg, 39.42 μmol, 59% yield).

Step 2: 1. To a solution of 1-1 (23.4 mg, 39.42 μmol) in THF (1 mL), 2mL of Ethanol (2 mL) and Methanol (1 mL) at ambient temperature wasadded aqueous LiOH (2 M, 2 mL). The mixture was stirred at 22° C. for 36hr. Cooled to −20° C. then quenched with aqueous HCl solution (2.0 M,2.1 mL) to acidic pH. The mixture was diluted with water (10 mL), andextracted with DCM (3×10 mL). The combined organic layer was washed withbrine and then dried over Na₂SO₄. Salts were filtered and the filtratewas concentrated under reduced pressure and dried under high vacuum. Thecrude was used directly without further purification and assumingquantitative yield. 2. Crude was dissolved in anhydrous DCM (2 mL) andHCl/4 M DIOXANE (4 M, 2 mL) was added. The mixture was stirred for 45min at 22° C., then concentrated to dryness under reduced pressurefollowed by high vacuum treatment. The obtained crude was used directlywithout further purification assuming quantitative yield. 3. AnhydrousDCM (1.90 mL) was added to the crude followed by Hunig's base (50.93 mg,394.05 μmol, 68.64 uL) and FDPP (22.71 mg, 59.11 μmol) in one portion.The reaction mixture was allowed to stir for 18 hr then quenchedreaction with 2 M Na₂CO₃ solution (5 mL). Mixture was stirred for 5 minthen extracted with DCM (3×10 mL). Combined organic extracts were driedwith Na₂SO₄ and concentrated under reduced pressure. Flashchromatography (ISCO system, silica 12 g, 0-10% methanol indichloromethane) provided compound 1 (10.8 mg, 24.14 μmol, 61.26% yield,3 steps).

Compounds 2 through 8 were prepared according to General Method F withstarting materials A-2 through A-8, respectively, with compound B.

MS [M + H] Cpd Structure m/z ¹H NMR (DMSO-d₆) δ ppm 1

448.17 (500 MHz) 9.14 (s, 1 H) 8.66 (s, 1 H) 8.00 (s, 1 H) 7.49 (dd, J =9.16, 2.86 Hz, 1 H) 7.00- 7.16 (m, 2 H) 6.36-6.64 (m, 1 H) 5.60 (d, J =14.89 Hz, 1 H) 5.04 (br t, J = 10.02 Hz, 1 H) 4.67 (d, J = 12.03 Hz, 1H) 4.34-4.41 (m, 1 H) 4.30 (d, J = 14.89 Hz, 1 H) 4.09 (d, J = 9.74 Hz,1 H) 3.88 (d, J = 9.74 Hz, 1 H) 1.63 (s, 3 H) 1.48 (s, 3 H) 2

460.17 (500 MHz) 8.65 (s, 1 H) 8.40-8.48 (m, 1 H) 8.06 (s, 1 H) 7.36 (brd, J = 9.16 Hz, 1 H) 6.98- 7.07 (m, 2 H) 6.50 (td, J = 54.70, 4.01 Hz, 1H) 5.71 (br d, J = 14.89 Hz, 1 H) 4.93-5.03 (m, 1 H) 4.78 (d, J = 10.31Hz, 1 H) 4.66 (d, J = 12.03 Hz, 1 H) 4.34-4.43 (m, 1 H) 4.25 (d, J =15.47 Hz, 1 H) 3.76-3.83 (m, 1 H) 3.46 (br d, J = 10.31 Hz, 1 H) 2.82(br dd, J = 13.75, 2.86 Hz, 1 H) 0.81-0.89 (m, 1 H) 0.65-0.76 (m, 3 H) 3

434.19 (500 MHz) 8.65 (s, 1 H) 8.23 (br d, J = 4.01 Hz, 1 H) 8.06 (s, 1H) 7.32 (br d, J = 8.59 Hz, 1 H) 7.02-7.06 (m, 2 H) 6.35-6.61 (m, 1 H)5.62 (d, J = 14.32 Hz, 1 H) 4.91-5.00 (m, 1 H) 4.67 (d, J = 12.03 Hz, 1H) 4.40 (br dd, J = 10.88, 5.16 Hz, 2 H) 4.20-4.29 (m, 2 H) 3.62-3.71(m, 1 H) 3.37-3.43 (m, 1 H) 2.03- 2.16 (m, 2 H) 4

446.09 (500 MHz) 9.09 (s, 1 H) 8.66 (s, 1 H) 7.99 (s, 1 H) 7.40 (dd, J =9.45, 3.15 Hz, 1 H) 7.04 (td, J = 8.45, 3.15 Hz, 1 H) 6.92-6.98 (m, 1 H)6.50 (td, J = 54.84, 3.72 Hz, 1 H) 5.62 (d, J = 14.32 Hz, 1 H) 4.96-5.04(m, 1 H) 4.68 (d, J = 12.03 Hz, 1 H) 4.32-4.38 (m, 1 H) 4.21- 4.31 (m, 2H) 3.91 (d, J = 10.31 Hz, 1 H) 1.89- 1.96 (m, 1 H) 0.95-1.07 (m, 2 H)0.73-0.80 (m, 1 H) 5

448.20 (300 MHz) 8.63 (s, 1 H) 8.21 (d, J = 6.51 Hz, 1 H) 8.05 (s, 1 H)7.31 (dd, J = 9.17, 2.75 Hz, 1 H) 6.92-7.07 (m, 2 H) 6.26-6.68 (m, 1 H)5.62 (d, J = 14.76 Hz, 1 H) 4.89-5.02 (m, 1 H) 4.66 (d, J = 12.10 Hz, 1H) 4.35 (br dd, J = 11.83, 2.75 Hz, 1 H) 4.08-4.28 (m, 4 H) 2.16-2.30(m, 1 H) 1.87-2.02 (m, 1 H) 1.26 (d, J = 6.24 Hz, 3 H) 6

460.20 (500 MHz) 8.62 (s, 1 H) 8.18 (s, 1 H) 7.96 (s, 1 H) 7.24 (dd, J =9.16, 2.86 Hz, 1 H) 7.07 (dd, J = 9.16, 4.58 Hz, 1 H) 6.96-7.03 (m, 1 H)6.49 (td, J = 54.56, 3.72 Hz, 1 H) 5.58 (br d, J = 14.89 Hz, 1 H)4.88-4.95 (m, 1 H) 4.66 (d, J = 12.03 Hz, 1 H) 4.36-4.54 (m, 2 H) 4.20-4.34 (m, 2 H) 2.09-2.22 (m, 1 H) 1.57-1.75 (m, 2 H) 1.12-1.17 (m, 1 H)0.75-0.84 (m, 1 H) 0.66-0.73 (m, 1 H) 7

462.17 (300 MHz) 8.64 (s, 1 H) 8.46 (br d, J = 3.21 Hz, 1 H) 8.07 (s, 1H) 7.40 (dd, J = 9.31, 2.89 Hz, 1 H) 7.01-7.17 (m, 2 H) 6.27-6.70 (m, 1H) 5.66 (d, J = 15.22 Hz, 1 H) 4.92-5.07 (m, 1 H) 4.66 (d, J = 12.10 Hz,1 H) 4.23-4.42 (m, 3 H) 3.60-3.68 (m, 1 H) 3.35-3.42 (m, 1 H) 3.22 (s, 1H) 1.26 (s, 3 H) 1.04 (s, 3 H) 8

462.22 (500 MHz) 8.61 (s, 1 H) 7.98-8.08 (m, 2 H) 7.41 (dd, J = 9.16,2.86 Hz, 1 H) 7.02-7.14 (m, 2 H) 6.46 (td, J = 54.56, 3.72 Hz, 1 H) 5.70(br d, J = 14.89 Hz, 1 H) 5.01 (br d, J = 12.60 Hz, 1 H) 4.64 (d, J =12.03 Hz, 1 H) 4.49 (br t, J = 10.02 Hz, 1 H) 4.29 (br d, J = 10.31 Hz,1 H) 4.21 (br d, J = 14.89 Hz, 1 H) 3.95 (br dd, J = 9.45, 5.44 Hz, 1 H)2.25 (br dd, J = 15.46, 9.74 Hz, 1 H) 1.87 (br dd, J = 15.46, 5.15 Hz, 1H) 1.59 (s, 3 H) 1.52 (s, 3 H)

Example 7: Biologic Assays In-Vitro Assays Materials and MethodsBiochemical Kinase Assay Method

The biochemical kinase assay was performed at Reaction BiologyCorporation (Malvern, PA) following the procedures described in thereference (Anastassiadis T, et al Nat Biotechnol. 2011, 29, 1039).Specific kinase/substrate pairs along with required cofactors wereprepared in reaction buffer; 20 mM Hepes pH 7.5, 10 mM MgCl₂, 1 mM EGTA,0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na₃VO₄, 2 mM DTT, 1% DMSO.Compounds were delivered into the reaction, followed about 20 min laterby addition of a mixture of ATP (Sigma, St. Louis MO) and ³³P ATP(Perkin Elmer, Waltham MA) to a final concentration of 10 μM. Reactionswere carried out at room temperature for 120 min, followed by spottingof the reactions onto P81 ion exchange filter paper (Whatman Inc.,Piscataway, NJ). Unbound phosphate was removed by extensive washing offilters in 0.75% phosphoric acid. After subtraction of backgroundderived from control reactions containing inactive enzyme, kinaseactivity data was expressed as the percent remaining kinase activity intest samples compared to vehicle (dimethyl sulfoxide) reactions. IC₅₀values and curve fits were obtained using Prism (GraphPad Software).

Cell Lines and Cell Culture:

Colorectal cell line KM 12 (harboring endogenous TPM3-TRKA fusion gene)was obtained from NCI. Acute myelogenous cell line KG-1 (harboringendogenous OP2-FGFR1 fusion gene) was purchased from ATCC.

Cloning and Ba/F3 Stable Cell Lines Creation

The EML4-ALK gene (variant 1) wild type, G1202R, G1202R/L1196M,G1202R/L1198F, G1202R/C1156Y, and L1196M/L1198F were synthesized atGenScript and cloned into pCDH-CMV-MCS-EF1-Puro plasmid (SystemBiosciences, Inc). Ba/F3 EML4-ALK wild type, G1202R, G1202R/L1196M,G1202R/L1198F, G1202R/C1156Y, L1196M/L1198F, L1198F/C1156Y,G1202R/G1269A, and G1202R/G1269A/L1204V were generated by transducingBa/F3 cells with lentivirus containing EML4-ALK wide type, G1202R,G1202R/L1196M, G1202R/L1198F, G1202R/C1156Y, L1196M/L1198FL1198F/C1156Y, G1202R/G1269A, or G1202R/G1269A/L1204V. Stable cell lineswere selected by puromycin treatment, followed by IL-3 withdrawal.Briefly, 5×10⁶ Ba/F3 cells were transduced with lentivirus supernatantin the presence of 8 μg/mL protamine sulfate. The transduced cells weresubsequently selected with 1 pg/mL puromycin in the presence ofIL3-containing medium RPMI1640, plus 10% FBS. After 10-12 days ofselection, the surviving cells were further selected for IL3 independentgrowth.

Cell Proliferation Assays:

-   -   Two thousand cells per well were seeded in 384 well white plate        for 24 hrs, and then treated with compounds for 72 hr (37° C.,        5% CO₂). Cell proliferation was measured using CellTiter-Glo        luciferase-based ATP detection assay (Promega) following the        manufactures's protocol. IC₅₀ determinations were performed        using GraphPad Prism software (GraphPad, Inc., San Diego, CA).

Immunoblotting for Cellular Kinase Phosphorylation Assays

Half a million cells (Ba/F3 EML4-ALK WT or G1202R) per well were seededin 24 well plate for 24 hr, and then treated with compound for 4 hr.Cells were collected after treatment and lysed in RIPA buffer (50 mMTris, pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% Deoxycholate, 0.1% SDS)supplemented with 10 mM EDTA, 1×Halt protease and phosphatase inhibitors(Thermo Scientific). Protein lysates (approximately 20 pg) was resolvedon 4-12% Bolt Bis-Tris precasted gels with MES running buffer (LifeTechnologies), transferred to nitrocellulose membranes using Trans-BlotTurbo Transfer System (Bio-Rad) and detected with antibodies targetingphosphorylated ALK (Y1282/1283) (Cell Signaling Technology), ALK(Y1604), total ALK (Cell Signaling Technology), Actin (Cell SignalingTechnology). Antibodies were typically incubated overnight at 4° C. withgentle shake, followed by washes and incubation with the appropriateHRP-conjugated secondary antibodies. Membranes were incubated withchemiluminescent substrate for 5 min at room temperature (SuperSignalWest Femto, Thermo Scientific). The chemiluminescent images wereacquired with a C-DiGit Imaging System (LI-COR Biosciences). Therelative density of the chemiluminescent bands was quantified via ImageStudio Digits from LICOR.

Solubility Test

Aliquot of 1.035 mL phosphate buffer (PB) (pH 7.4) was added into 2.07mg of the test compound (for a 2 mg mL⁻¹ upper limiting solubilitydetermination) in a 1.5 mL tubes. The mixture was ultrasonicly treatedfor 10 min and rotated on rotator at room temperature for more than 8hr. After rotation, the tube was ultrasonicly treated for 10 min andcentrifuged at 13000 rpm for 15 min. 0.3 mL of the supernatant wastransferred into a 0.6 mL tube and discarded after rotated for 5 min forrinsing, then about 0.6 mL of the residual supernatant was transferredinto the rinsed tube and centrifuged again at 13000 rpm for 15 min. Thesupernatant after every centrifugation was appropriately diluted andtransferred for LC-MS/MS analysis where a standard curve was constructedwith 7 concentration levels (2, 4, 10, 20, 40, 100 and 200 g mL⁻¹) andused for the quantitation of the analyte in PB.

Liver Microsomal Stability Studies

1. Preparation of Stock and Working Solution

The test compound stock solution was prepared by weighing 1.07 mg anddissolving in 0.261 mL of DMSO to achieve the concentration of 10 mM.Working solution of the test compound was prepared by diluting stocksolution using DMSO to yield the concentration of 300 μM.

2. Incubation

The assay was carried out in 96-well microtiter plates. In each well,the reaction mixture (25 μL) contained the test compound at the finalconcentration of 1 μM, 0.5 mg/mL liver microsome protein, and 1 mM NADPHin 100 mM potassium phosphate, pH 7.4 buffer with 3.3 mM MgCl₂. Themixtures were incubated in duplicate at 37° C. for 0, 15, 30 or 60minutes, and 150 μL of quench solution (acetonitrile with 0.1% formicacid) with internal standard (bucetin for positive ESI mode) was addedinto each well of reaction to terminate the reaction. The plates werethen sealed and centrifuged at 4° C. for 15 minutes at 4000 rpm. Theresulting supernatant was transferred to fresh plates for LC-MS/MSanalysis of the test compounds. Verapamil was used as the positivecontrol to validate the assay system.

3. LC-MS/MS Analysis

All samples were analyzed with LC-MS/MS using an AB Sciex API 4000instrument, coupled to a Shimadzu LC-20AD LC Pump system. Analyticalsamples were separated using a Waters Atlantis T3 dC18 reverse phaseHPLC column (20 mm×2.1 mm) at a flow rate of 0.5 mL/min. The mobilephase consisted of 0.1% formic acid in water (solvent A) and 0.1% formicacid in acetonitrile (solvent B).

4. Calculation

The extent of metabolism was calculated based on the disappearance ofthe test compound, compared to its initial concentration. The initialrates of clearance of test compounds were calculated using linearregression plot of semi-log % remaining of compounds versus time. Theelimination rate constant (equals to negative slope) of the linearregression plot was then used to determine t_(1/2) and the intrinsicclearance (CL_(int)) using the following formula:

-   -   k=−slope    -   t_(1/2)=0.693/k    -   CL_(int)=k/C_(protein)

Where C_(protein) (mg/mL) is the microsomal protein concentration in theincubation system. This method of intrinsic clearance determinationmakes the assumption that the test compound concentration is far belowthe Michaelis-Menten constant of the compound to its metabolizingenzymes.

In-Vivo Studies Subcutaneous Xenograft Models in Immune Compromised Mice

Female SCID/Beige mice (5-8 weeks of age) were obtained from CharlesRiver Laboratory and were housed in Innovive IVC disposable cages onHEPA filtered ventilated racks with ad libitum access to rodent chow andwater. About five million cells in 100 μL serum-free medium supplementedwith 50% matrigel (Corning, Inc) were implanted subcutaneously in theright flank region of the mouse. Tumor size and body weight weremeasured on designated days. Tumor size was measured with an electroniccaliper and tumor volume was calculated as the product oflength*width²*0.5. Mice were randomized by tumor size into treatmentgroups when tumor volume reached about 100-200 mm³. Compound 1 or 2 wasadministered orally at pre-determined schemes and doses. Vehicle wasused as negative control. Lorlatinib was used as a reference forefficacy evaluation. Tumor growth inhibition (TGI) was calculated asfollows: If TV_(t)>TV₀, TGI=100%×(1−(TV_(t)−TV₀)/(CV_(t)−TV₀)); IfTV_(t)<TV₀, TGI=100%×(2−TV_(t)/TV₀); where TV₀ was the mean tumor volumein the treatment group at the beginning of the treatment, TV_(t) was themean tumor volume in the treatment group at the end of the treatment,CV₀ was the mean tumor volume in the vehicle control group at thebeginning of the treatment, and CV_(t) was the mean tumor volume in thevehicle control group at the end of the treatment. A TGI that is largerthan 100% indicates tumor regression. Statistical analyses wereperformed using GraphPad Prism 8.4.0 and p<0.05 was considered asstatistically significant difference.

Tumor Processing and Immunoblotting for In Vivo Pharmacodynamic Studies

Mice bearing xenograft tumors were humanely euthanized and tumors wereresected and snap frozen in liquid nitrogen and stored at −80° C. Frozentumor samples were processed at 4° C. in 1× Cell Lysis Buffer (CellSignaling Technologies) to extract proteins. SDS loading samples wereprepared by addition of 4×LDS Sample Buffer and 10× Reducing Reagent(Life Technologies, Inc) to protein lysate. Tumor SDS protein sampleswere processed by SDS-PAGE and immunoblotted with rabbitanti-phosphorylated ALK Y1282/1283, rabbit anti-phosphorylated ALKY1604, rabbit anti-ALK and mouse anti-actin antibodies (Cell SignalingTechnologies). The signals from immunoblot were detected by C-DiGit BlotScanner from LI-COR and the signal intensity were quantified using theImage Studio Digit software (LI-COR).

Mouse Pharmacokinetic Studies

Preparation of the Vehicle Solution for the Test Article

To prepare 1 L of the vehicle solution (0.5% CMC and 1.0% Tween 80), 10g of Tween 80 was mixed with 985 mL of water into a 1 L bottle. Themixture was stirred until Tween 80 was completely dissolved. Withcontinued stirring, 5 g of CMC was very slowly sprinkled into thesolution. Stirring was continued until all CMC was dissolved, whichmight take several hr. The resulting vehicle solution was stored at 4°C.

PK Blood Sample Processing and Bioanalysis Procedure

The compound was suspended in the vehicle solution and the mice wereorally administrated with a single dose of the compound at the selecteddose level. The blood samples were collected according to the definedtime table into tubes containing K2-EDTA, followed by gentle mixing toassure distribution of the anti-coagulant. Immediately after the bloodsample was collected and mixed, it was placed on ice. The samples werethen centrifuged at 4° C. for 10 minutes at 5,000 RPM. The plasma washarvested into pre-labeled tubes and stored at −80° C. until analyzed byLC-MS/MS.

In-Vitro Results Enzymatic Kinase Activities Against ALK and Mutant ALKs

Compounds were tested against ALK and mutant ALKs in the enzymatickinase catalytic activity assays at Reaction Biology Corporation. Theresults were reported in Table 1. Compound 1 (“Cpd 1”) and Compound 2(“Cpd 2”) each showed potent kinase inhibitory activities on ALK andmutant ALKs with IC₅₀s<10 nM.

TABLE 1 Kinase Cpd 1 IC₅₀ (nM) Cpd 2 IC₅₀ (nM) ALK 1.43 1.40 ALK(C1156Y) 0.19 0.11 ALK (D1203N) 4.36 2.90 ALK (delete G1202) 0.53 0.45ALK (E1210K) 0.34 0.21 ALK (E1210K/D1203N) 6.29 4.00 ALK (E1210K/S1206C)0.22 0.13 ALK (F1174C) 1.78 0.92 ALK (F1174L) 0.69 0.42 ALK (F1174S)1.20 0.76 ALK (F1245C) 0.75 0.41 ALK (G1202R) 0.88 0.74 ALK (G1269A)1.55 1.28 ALK (G1269S) 6.60 6.14 ALK (I117IN) 2.29 1.16 ALK (L1152P)2.92 1.96 ALK (L1152R) 1.10 0.83 ALK (L1196M) 0.29 0.18 ALK (L1198F)1.05 0.85 ALK (L1198F/C1156Y) 0.24 0.15 ALK (L1198F/G1202R) 0.63 0.49ALK (L1198F/L1196M) 0.18 0.10 ALK (R1275Q) 0.82 0.72 ALK (S1206R) 0.500.31 ALK (T1151-L1152insT) 1.25 0.86 ALK (T1151M) 0.39 0.28 ALK (V1180L)1.61 0.51

Anti-Cell Proliferation Activity

Compounds were tested in cell proliferation assays in Ba/F3 cellsengineered with EML4-ALK and EML4-ALK G1202R, and also in KM 12 cellshaving TPM3-TRKA fusion and KG-1 cells having OP2-FGFR1 fusion. Theresults were summarized in Table 2.

TABLE 2 BaF3 BaF3 EML4-ALK KM12 KG-1 cell EML4-ALK G1202R (TPM3-TRKA)(OP2-FGFR1) Cpd IC₅₀ (nM) IC₅₀ (nM) IC50 (nM) IC₅₀ (nM) 1 0.8 0.2 <0.2554.2 2 <0.2 0.2 <0.2 3.8 3 9.7 9.3 <0.2 75.3 4 9 35.7 N/A 60.9 5 46.3NA <0.2 513.5 6 27.7 152.9 <0.2 385.3 7 22 NA <0.2 40.7 8 186 122.5 <0.22095

Comparative Examples 1 and 2 were also evaluated for anti-cellproliferation activity. PP-1T The results were summarized in Table 2-A.

TABLE 2-A BaF3 BaF3 KM12 KG-1 cell EML4-ALK EML4-ALK G1202R (TPM3-TRKA)(OP2-FGFR1) Cpd IC₅₀ (nM) IC₅₀ (nM) IC₅₀ (nM) IC₅₀ (nM) Comp. Ex. 1 22.843.0 <0.2 22.2 Comp. Ex. 2 <0.6 <1 0.2 0.65 BaF3 BaF3 BaF3 BaF3 BaF3JAK2 V617F KIF5B-RET KIF5B-RET G810R KIF5B-RET V804M TEL-TRKB Cpd IC₅₀(nM) IC₅₀ (nM) IC₅₀ (nM) IC₅₀ (nM) IC₅₀ (nM) Comp. Ex. 1 825 110 787 1380.74 Comp. Ex. 2 3450 180 1440 490 <0.2Anti-Cell Proliferation in Ba/F3 Cells Engineered with Mutant ALKs

Compounds were tested in cell proliferation assays in Ba/F3 cellsengineered with EML4-ALK wildtype (WT) and mutant ALKs. The results weresummarized in Table 3 and 3-A.

TABLE 3 Ba/F3 Cell Proliferation IC₅₀ (nM) ALK ALK ALK ALK ALK ALK ALKG1202R/ ALK ALK G1202R/ G1202R/ G1202R/ L1196M/ L1198F/ G1202R/ G1269A/Cpd WT G1202R L1196M L1198F C1156Y L1198F C1156Y G1269A L1204V 1 0.8 0.21.1 <0.2 <0.2 <0.2 <0.2 8.7 14.7 2 <0.2 0.2 0.5 <0.2 <0.2 <0.2

A side-by-side comparison of Compound 1 with known kinase inhibitors inBa/F3 cells engineered with EML4-ALK wildtype (WT) and mutant ALKs isshown in Table 3-A.

TABLE 3-A Ba/F3 Cell proliferation IC₅₀ (nM) EML4-ALK Compound 1Crizotinib Alectinib Brigatinib Ceritinib Lorlatinib WT 0.4 50 7.4 123.9 0.8 I1171N 516 254 4310 49 72 48 I1171S 189 188 306 31 27 31 I1171T316 232 210 33 29 25 L1196M 0.5 274 50.1 21.1 5.4 38.2 L1198F <0.2 18397 74 618 30 G1202R 0.2 434 2690 188 329 52 G1269A 13 451 197 20 15 49G1269S 701 1390 671 46 97 191 L1196M/L1198F <0.2 252 2250 253 1410 1310L1198F/C1156Y <0.2 19.3 776 102 1310 140 L1198F/I1171N 1.6 626 236 55.164.1 78.7 G1202R/C1156Y 0.2 745 2420 810 1300 521 G1202R/L1196M 0.7808 >10000 1100 1260 4780 G1202R/L1198F <0.2 188 3000 2040 2010 1710G1202R/G1269A 9.9 705 7200 164 303 636 G1202R/G1269A/L1204V 14.9 6346740 176 345 673 G1202R/G1269A/L1198F 0.2 596 >10000 907 1670 6330

Solubility

The solubility was tested at pH 7.4 and the results were summarized inTable 4.

TABLE 4 Solubility (μg/mL) Compound at pH 7.4 Comp. Ex. 1 7.9 Comp. Ex.2 4.9 Compound 1 3.3 Compound 2 2.6

Liver Microsomal Stability

The liver microsomal stability was tested and the results weresummarized in Table 5.

TABLE 5 Microsomal Stability Cl_(int) (μL/min/mg) Human Mouse RatCompound (pooled) (female) (female) Comp. Ex. 1 2.6 22 10 Comp. Ex. 23.9 21 5.9 Compound 1 8.3 11 1.5 Compound 2 8.8 25 3.5

In-Vivo Results Mouse PK

The mouse PKs were determined and the results were summarized in Table6.

TABLE 6 Compound C_(max) (ng/mL) T_(max) (h) AUC_(all) (h*ng/mL) Comp.Ex. 1 4190 0.5 12300 Comp. Ex. 2 2900 0.5 18700 Compound 1 1751 2.011900 Compound 2 2510 2.0 14600Effect of Compound 1 and 2 on Ba F3 Cell-Derived Xenograft Tumors withthe EML4-ALK G1202R Fusion

SCID/Beige mice bearing Ba/F3 cell-derived tumors with the EML4-ALKG1202R fusion were randomized to seven groups with 8 mice per group onday 8 post inoculation and treated with vehicle BID, Compound 1 BID at 2mg/kg, Compound 1 BID at 5 mg/kg, Compound 1 BID at 10 mg/kg, Compound 2BID at 3 mg/kg, Compound 2 BID at 10 mg/kg and lorlatinib BID at 5mg/kg, respectively. The tumor volume vs time data are shown as mean±semin FIG. 1A. After 7 days of treatment, Compound 1 treatment inhibitedtumor growth or led to tumor regression with TGIs of 64%, 120% and 200%at dose of 2, 5, and 10 mg/kg BID, respectively; in comparison,lorlatinib inhibited tumor growth with a TGI of 154% at 5 mg/kg BID. Thestatistical evaluation of Compound 1's effect on tumor volume includedthe vehicle treated group, Compound 1 treated groups at 2, 5, 10 mg/kgBID and the lorlatinib treated group, using mixed-effects model followedby post hoc Tukey's multiple comparisons test. The efficacy of Compound1 on this model with EML4-ALK G1202R fusion started to be detectable onday 10 and the tumor volume in the groups treated with Compound 1 at allthree dose levels is significantly smaller than that of the grouptreated with vehicle by the last day of treatment (on day 15, p=0.0007for Compound 1 2 mg/kg vs vehicle and p<0.0001 for Compound 1 5 or 10mg/kg vs vehicle). Compound 1 treatment at 10 mg/kg BID is moreeffective than treatment by lorlatinib at 5 mg/kg BID in this model withthe EML4-ALK G1202R fusion, as indicated by significantly smaller tumorvolume in the group treated with Compound 1 at 10 mg/kg BID than that ofthe group treated with lorlatinib on days 13, 14, 15 post inoculation(p<0.0001 on day 13 and 14, p=0.0003 on day 15).

The body weight vs time data are shown as mean±sem in FIG. 1B. There wasno body weight loss in groups treated with Compound 1 at 2, 5, 10 mg/kgBID after 7 days of treatment. The statistical evaluation of Compound1's effect on body weight included vehicle the treated group, Compound 1treated groups at 2, 5, 10 mg/kg BID and the lorlatinib treated group,using mixed-effects model followed by post hoc Tukey's multiplecomparisons test. The body weight in the group treated with Compound 1at 2 mg/kg BID was significantly higher than that of the group treatedwith vehicle on days 13, 14, 15 post inoculation (p=0.0012 on day 13,p=0.0001 on day 14, p<0.0001 on day 15). The body weight in the grouptreated with Compound 1 at 5 mg/kg BID was significantly higher thanthat of the group treated with vehicle on days 13, 14, 15 postinoculation (p=0.0392 on day 13, p=0.0126 on day 14, p=0.0048 on day15). The body weight in the group treated with Compound 1 at 10 mg/kgBID was significantly higher than that of the group treated with vehicleon days 13, 14 post inoculation (p=0.0495 on day 13, p=0.0250 on day14).

After 7 days of treatment, Compound 2 treatment inhibited tumor growthwith TGIs of 51% and 77% at dose of 3 and 10 mg/kg BID, respectively;whereas lorlatinib inhibited tumor growth with a TGI of 154% at 5 mg/kgBID. The statistical evaluation Compound 2's effect on tumor volumeincluded the vehicle treated group, Compound 2 treated groups at 3 and10 mg/kg BID and the lorlatinib treated group, using mixed-effects modelfollowed by post hoc Tukey's multiple comparisons test. The effect ofCompound 2 on tumor growth inhibition started to be detectable by day 13at the 3 mg/kg dose level (p=0.0019 vs vehicle) and by day 10 at the 10mg/kg dose level (p=0.0396 vs vehicle). On day 15, the last day oftreatment, the tumor volume in the group treated with Compound 2 wassignificantly smaller than that of the group treated with vehicle(p=0.0272 for Compound 2 3 mg/kg vs vehicle; p=0.0052 for Compound 2 10mg/kg vs vehicle).

There was no body weight loss in groups treated with Compound 2 at 3 and10 mg/kg BID after 7 days of treatment. The statistical evaluation ofCompound 2's effect on body weight included the vehicle treated group,Compound 2 treated groups at 3 and 10 mg/kg BID and the lorlatinibtreated group, using mixed-effects model followed by post hoc Tukey'smultiple comparisons test. The body weight in the group treated withCompound 2 at 3 mg/kg BID was significantly higher than that of thegroup treated with vehicle on days 13, 14, 15 post inoculation (p=0.0282on day 13, p=0.0229 on day 14, p=0.0043 on day 15). The body weight inthe group treated with Compound 2 at 10 mg/kg BID was significantlyhigher than that of the group treated with vehicle on days 13, 14, 15post inoculation (p=0.0017 on day 13, p=0.0009 on day 14; p=0.0003 onday 15).

Effect of Compound 1 on Ba/F3 Cell-Derived Xenograft Tumors with theEML4-ALK L1198F/G1202R Fusion

SCID/Beige mice bearing Ba/F3 cell-derived tumors with the EML4-ALKL1198F/G1202R fusion were randomized to five groups with 10 mice pergroup on day 7 post inoculation and treated with vehicle BID, Compound 1BID at 2 mg/kg, Compound 1 BID at 5 mg/kg, Compound 1 BID at 10 mg/kg,and lorlatinib BID at 5 mg/kg, respectively. The tumor volume vs timedata are shown as mean±sem in FIG. 2A. After 7 days of treatment,Compound 1 treatment led to complete tumor regression with TGIs of 200%at all three dose levels of 2, 5, and 10 mg/kg BID. Tumors in all threegroups treated with Compound 1 exhibited complete regression at the endof study on day 14 post inoculation. The inhibitory activity of Compound1 in this model with the EML4-ALK L1198F/G1202R fusion was much betterthan that of lorlatinib, which inhibited tumor growth to a much lessextent with a TGI of 310% at 5 mg/kg BID after 7 days of treatment. Thestatistical evaluation of Compound 1's effect on tumor volume wasperformed using two-way repeated measures ANOVA followed by post hocTukey's multiple comparisons test. The tumor volume in the group treatedwith Compound 1 at all three dose levels was significantly smaller thanthat of the group treated with vehicle on days 10, 12, 13, 14 postinoculation (p<0.0001 for Compound 1 2, 5, or 10 mg/kg vs vehicle oneither day 10, 12, 13 or 14). Furthermore, the tumor volume in the grouptreated with Compound 1 at all three dose levels was significantlysmaller than that of the group treated with lorlatinib on days 10, 12,13, 14 post inoculation (p<0.0001 for Compound 1 2, 5, or 10 mg/kg vslorlatinib on either day 10, 12, 13 or 14).

The body weight vs time data are shown as mean±sem in FIG. 2B. There wasno body weight loss in groups treated with Compound 1 at 2, 5, 10 mg/kgBID after 7 days of treatment. The statistical evaluation of Compound1's effect on body weight was performed using two-way repeated measuresANOVA followed by post hoc Tukey's multiple comparisons test. The bodyweight in the group treated with Compound 1 at 2 mg/kg BID wassignificantly higher than that of the group treated with vehicle on days10, 12, 13, 14 post inoculation (p=0.0478 on day 10, p=0.0044 on day 12,p=0.0008 on day 13, p=0.0013 on day 14). The body weight in the grouptreated with Compound 1 at 5 mg/kg BID was significantly higher thanthat of the group treated with vehicle on days 12, 13, 14 postinoculation (p=0.0308 on day 12, p=0.0056 on day 13, p=0.0050 on day14). The body weight in the group treated with Compound 1 at 10 mg/kgBID was significantly higher than that of the group treated with vehicleon days 10, 12, 13 post inoculation (p=0.0390 on day 10, p=0.0048 on day12, p=0.0119 on day 13).

Effect of Compound 1 on Ba/F3 Cell-Derived Xenograft Tumors with theEML4-ALK L1196M/G1202R Fusion

SCID/Beige mice bearing Ba/F3 cell-derived tumors with the EML4-ALKL1196M/G1202R fusion were randomized to six groups with 8 mice per groupon day 7 post inoculation and treated with vehicle BID, Compound 1 BIDat 0.6 mg/kg, Compound 1 BID at 2 mg/kg, Compound 1 BID at 5 mg/kg,Compound 1 BID at 10 mg/kg, and lorlatinib BID at 5 mg/kg, respectively.The tumor volume vs time data are shown as mean±sem in FIG. 3A. After 7days of treatment, Compound 1 treatment resulted in TGIs of 19%, 44%,83% and 200% at dose levels of 0.6, 2, 5, and 10 mg/kg BID,respectively; whereas lorlatinib inhibited tumor growth with a TGI of18% at 5 mg/kg BID after 7 days of treatment. The statistical evaluationof Compound 1's effect on tumor volume was performed using mixed-effectsmodel followed by post hoc Tukey's multiple comparisons test. The tumorgrowth in the groups treated with Compound 1 at 0.6 mg/kg BID orlorlatinib at 5 mg/kg BID was not significantly different from vehicletreated group after 7 days of treatment (p>0.05 for vehicle vslorlatinib or Compound 1 0.6 mg/kg on day 14). Compound 1 at 2, 5 and 10mg/kg BID exhibited effectiveness in this model with the EML4-ALKL1196M/G1202R fusion. After seven days of treatment, the tumor volume inthe group treated with Compound 1 at 2, 5 or 10 mg/kg BID wassignificantly smaller than that of the group treated with vehicle (onday 14, p=0.0013 for Compound 1 2 mg/kg vs vehicle, p=0.0001 forCompound 1 5 mg/kg vs vehicle, p<0.0001 for Compound 1 10 mg/kg vsvehicle). Moreover, after seven days of treatment, the tumor volume inthe group treated with Compound 1 at 2, 5 or 10 mg/kg BID wassignificantly smaller than that of the group treated with lorlatinib (onday 14, p=0.0070 for Compound 1 2 mg/kg vs lorlatinib, p=0.0018 forCompound 1 5 mg/kg vs lorlatinib, p<0.0001 for Compound 1 10 mg/kg vslorlatinib).

The body weight vs time data are shown as mean±sem in FIG. 3B. There wasno body weight loss in groups treated with Compound 1 at 2, 5, 10 mg/kgBID after 7 days of treatment. The statistical evaluation of Compound1's effect on body weight was performed using mixed-effects modelfollowed by post hoc Tukey's multiple comparisons test. The body weightin the group treated with Compound 1 at 0.6 mg/kg BID was significantlyhigher than that of the group treated with vehicle on days 12, 13, 14post inoculation (p=0.0005 on day 12, p=0.0163 on day 13, p=0.0011 onday 14). The body weight in the group treated with Compound 1 at 2 mg/kgBID was significantly higher than that of the group treated with vehicleon day 14 post inoculation (p=0.0391 on day 14). The body weight in thegroup treated with Compound 1 at 5 mg/kg BID was significantly higherthan that of the group treated with vehicle on days 10, 12, 13, 14 postinoculation (p=0.0413 on day 10, p=0.0018 on day 12, p<0.0001 on day 13,p=0.0011 on day 14). The body weight in the group treated with Compound1 at 10 mg/kg BID was not significantly different from that of the grouptreated with vehicle (p>0.05 on day 10, 12, 13 and 14).

Effect of Compound 1 on Phosphorylation of ALK Fusion Protein in Ba/F3Cell-Derived Xenograft Tumors with the EML4-ALK L1196M/G1202R Fusion

The pharmacodynamic effect of Compound 1 was evaluated in the Ba/F3cell-derived xenograft tumors with the EML4-ALK L1196M/G1202R fusionfollowing a single oral dose of Compound 1 at 10 mg/kg. At 2 hours postdose, tumor samples were collected from mice treated with either vehicleor Compound 1 with three mice for each treatment. Samples were processedand analyzed by immunoblotting and the result is shown in FIG. 4 . Thelevels of phosphorylation of ALK fusions at Y1282/1283 and at Y1604 wereboth reduced in samples from the Compound 1 treated mice compared tothose from vehicle treated mice. The level of ALK fusion expression wasalso reduced in the samples from Compound 1 treated mice compared tothose from vehicle treated mice. As the control, the expression level ofactin was not affected by Compound 1 treatment.

FIG. 5 shows ALK phosphorylation level post-in vivo dosing of Compound 1at 0.6, 2, or 5 mg/kg after 2 hours or 12 hours. At each dose,phosphorylation level was significantly reduced compared to control at 2hours, but only showed reduction at 12 hours at the highest dose.

1. A compound of the formula I

wherein each R¹ and R² is independently H, deuterium, halogen, C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic heteroaryl, —OR^(a),—OC(O)R^(a), —OC(O)R^(a), —OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a),—SR^(a), —S(O)R^(a), —S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b),—OS(O)NR^(a)R^(b), —OS(O)₂NR^(a)R^(b), —NR^(a)R^(b), —NR^(a)C(O)R^(b),—NR^(a)C(O)OR^(b), —NR^(a)C(O)NR^(a)R^(b), —NR^(a)S(O)R^(b),—NR^(a)S(O)₂R^(b), —NR^(a)S(O)NR^(a)R^(b), —NR^(a)S(O)₂NR^(a)R^(b),—C(O)R^(a), —C(O)OR^(a), —C(O)NR^(a)R^(b), —PR^(a)R^(b),—P(O)R^(a)R^(b), —P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b),—P(O)OR^(a), —P(O)₂OR^(a), —CN, or —NO₂; or R¹ and R² taken togetherwith the carbon atom to which they are attached combine to form a C₃-C₆cycloalkyl; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl,C₆-C₁₀ aryl, and mono- or bicyclic heteroaryl is independentlyoptionally substituted by deuterium, halogen, C₁-C₆ alkyl, C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f),—OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f),—SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f),—NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂; each R³ and R^(3′) is independently H,deuterium, halogen, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- orbicyclic heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),—OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a), —S(O)R^(a),—S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b), —OS(O)NR^(a)R^(b),—OS(O)₂NR^(a)R^(b), —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b),—NR^(a)S(O)NR^(a)R^(b), —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a),—C(O)OR^(a), —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),—P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b), —P(O)OR^(a),—P(O)₂OR^(a), —CN, or —NO₂, wherein each hydrogen atom in C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic heteroaryl, isindependently optionally substituted by deuterium, halogen, C₁-C₆ alkyl,C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),—OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),—OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f),—S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂; or R³ and R^(3′) taken together with thecarbon atom to which they are attached combine to form a C₃-C₆cycloalkyl; wherein each hydrogen atom in C₃-C₆ cycloalkyl isindependently optionally substituted by deuterium, halogen, C₁-C₆ alkyl,C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),—OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),—OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f),—S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR^(f),—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂; each R⁴ and R⁵ is independently hydrogen,deuterium, halogen, —OR^(c), —OC(O)R^(c), —OC(O)NR^(c)R^(d),—OC(═N)NR^(c)R^(d), —OS(O)R^(c), —OS(O)₂R^(c), —OS(O)NR^(c)R^(d),—OS(O)₂NR^(c)R^(d), —SR^(c), —S(O)R^(c), —S(O)₂R^(c), —S(O)NR^(c)R^(d),—S(O)₂NR^(c)R^(d), —NR^(c)R^(d), —NR^(c)C(O)R^(d), —NR^(c)C(O)OR^(d),—NR^(c)C(O)NR^(c)R^(d), —NR^(c)C(═N)NR^(c)R^(d), —NR^(c)S(O)R^(d),—NR^(c)S(O)₂R^(d), —NR^(c)S(O)NR^(c)R^(d), —NR^(c)S(O)₂NR^(c)R^(d),—C(O)R^(c), —C(O)OR^(c), —C(O)NR^(c)R^(d), —C(═N)NR^(c)R^(d),—PR^(c)R^(d), —P(O)R^(c)R^(d), —P(O)₂R^(c)R^(d), —P(O)NR^(c)R^(d),—P(O)₂NR^(c)R^(d), —P(O)OR^(c), —P(O)₂OR^(c), —CN, —NO₂, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic heteroaryl, whereineach hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- orbicyclic heteroaryl, C₅-C₈ cycloalkyl, or 5- to 8-memberedheterocycloalkyl is independently optionally substituted by deuterium,halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),—OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e),—OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e),—S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f),—NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f),—NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f),—C(O)R^(e), —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f),—P(O)R^(e)R^(f), —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),—P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂; R⁶ is H, deuterium, or C₁-C₆alkyl, wherein each hydrogen atom in C₁-C₆ alkyl is independentlyoptionally substituted by deuterium, halogen, —OR^(e), —SR^(e), or—NR^(e)R^(f); each R⁷ is independently hydrogen or deuterium; eachR^(a), R^(b), R^(c), R^(d), R^(e), and R is independently selected fromthe group consisting of H, deuterium, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀aryl, and 5- to 7-membered heteroaryl; and n is 0, 1, 2, or 3; or apharmaceutically acceptable salt thereof.
 2. The compound of claim 1, ora pharmaceutically acceptable salt thereof, wherein n is 0 or
 1. 3. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein n is
 0. 4. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein n is
 1. 5. The compound of claim 1,having the formula II

or a pharmaceutically acceptable salt thereof.
 6. The compound of claim1, having the formula IIa

or a pharmaceutically acceptable salt thereof.
 7. The compound of claim1, having the formula III

or a pharmaceutically acceptable salt thereof.
 8. The compound of claim1, having the formula IIIa

or a pharmaceutically acceptable salt thereof.
 9. The compound of claim1, or a pharmaceutically acceptable salt thereof, wherein each R¹ andR², when present, is independently H, deuterium, C₁-C₆ alkyl, or R¹ andR² taken together with the carbon atom to which they are attachedcombine to form a C₃-C₆ cycloalkyl.
 10. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein each R³ and R^(3′) isindependently H, deuterium, C₁-C₆ alkyl, or R³ and R^(3′) taken togetherwith the carbon atom to which they are attached combine to form a C₃-C₆cycloalkyl.
 11. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein each R^(3′) is H or deuterium.
 12. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein each R⁴ is H or deuterium.
 13. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein R⁵ is fluoro.
 14. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein R⁶ is H.
 15. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein each R⁷ is H.
 16. The compound of claim1, selected from the group consisting of

or a pharmaceutically acceptable salt thereof.
 17. A pharmaceuticalcomposition comprising a compound of claim 1, or a pharmaceuticallyacceptable salt thereof, and optionally at least one diluent, carrier orexcipient.
 18. A method of treating cancer in a subject in need thereof,comprising administering to the subject an effective amount of acompound of the formula I

or pharmaceutically acceptable salt thereof, wherein each R¹ and R² isindependently H, deuterium, halogen, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀aryl, mono- or bicyclic heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),—OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a), —S(O)R^(a),—S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b), —OS(O)NR^(a)R^(b),—OS(O)₂NR^(a)R^(b), —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b),—NR^(a)S(O)NR^(a)R^(b), —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a),—C(O)OR^(a), —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),—P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b), —P(O)OR^(a),—P(O)₂OR^(a), —CN, or —NO₂ or R¹ and R² taken together with the carbonatom to which they are attached combine to form a C₃-C₆ cycloalkyl;wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, andmono- or bicyclic heteroaryl is independently optionally substituted bydeuterium, halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),—OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e),—OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e),—S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f),—NR^(e)C(O)OR, —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f),—NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f),—C(O)R^(e), —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f),—P(O)R^(e)R^(f), —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),—P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂; each R³ and R^(3′) isindependently H, deuterium, halogen, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀aryl, mono- or bicyclic heteroaryl, —OR^(a), —OC(O)R^(a), —OC(O)R^(a),—OC(O)NR^(a)R^(b), —OS(O)R^(a), —OS(O)₂R^(a), —SR^(a), —S(O)R^(a),—S(O)₂R^(a), —S(O)NR^(a)R^(b), —S(O)₂NR^(a)R^(b), —OS(O)NR^(a)R^(b),—OS(O)₂NR^(a)R^(b), —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)OR^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)S(O)R^(b), —NR^(a)S(O)₂R^(b),—NR^(a)S(O)NR^(a)R^(b), —NR^(a)S(O)₂NR^(a)R^(b), —C(O)R^(a),—C(O)OR^(a), —C(O)NR^(a)R^(b), —PR^(a)R^(b), —P(O)R^(a)R^(b),—P(O)₂R^(a)R^(b), —P(O)NR^(a)R^(b), —P(O)₂NR^(a)R^(b), —P(O)OR^(a),—P(O)₂OR^(a), —CN, or —NO₂, wherein each hydrogen atom in C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, mono- or bicyclic heteroaryl, isindependently optionally substituted by deuterium, halogen, C₁-C₆ alkyl,C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),—OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),—OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f),—S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R, —NR^(e)C(O)OR^(f),—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R, —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂; or R³ and R^(3′) taken together with thecarbon atom to which they are attached combine to form a C₃-C₆cycloalkyl: wherein each hydrogen atom in C₃-C₆ cycloalkyl isindependently optionally substituted by deuterium, halogen, C₁-C₆ alkyl,C₁-C₆haloalkyl, —OR^(e), —OC(O)R^(e), —OC(O)NR^(e)R^(f),—OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e), —OS(O)NR^(e)R^(f),—OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), —S(O)NR^(e)R^(f),—S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f), —NR^(e)C(O)OR,—NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f),—NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f), —C(O)R^(e),—C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f), —P(O)R^(e)R^(f),—P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f), —P(O)OR^(e),—P(O)₂OR^(e), —CN, or —NO₂; each R⁴ and R⁵ is independently hydrogen,deuterium, halogen, —OR^(c), —OC(O)R^(c), —OC(O)NR^(c)R^(d),—OC(═N)NR^(c)R^(d), —OS(O)R^(c), —OS(O)₂R^(c), —OS(O)NR^(c)R^(d),—OS(O)₂NR^(c)R^(d), —SR^(c), —S(O)R^(c), —S(O)₂R^(c), —S(O)NR^(c)R^(d),—S(O)₂NR^(c)R^(d), —NR^(c)R^(d), —NR^(c)C(O)R^(d), —NR^(c)C(O)OR^(d),—NR^(c)C(O)NR^(c)R^(d), —NR^(c)C(═N)NR^(c)R^(d), —NR^(c)S(O)R^(d),—NR^(c)S(O)₂R^(d), —NR^(c)S(O)NR^(c)R^(d), —NR^(c)S(O)₂NR^(c)R^(d),—C(O)R^(c), —C(O)OR^(c), —C(O)NR^(c)R^(d), —C(═N)NR^(c)R^(d),—PR^(c)R^(d), —P(O)R^(c)R^(d), —P(O)₂R^(c)R^(d), —P(O)NR^(c)R^(d),—P(O)₂NR^(c)R^(d), —P(O)OR^(c), —P(O)₂OR^(c), —CN, —NO₂, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic heteroaryl, whereineach hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, mono- orbicyclic heteroaryl, C₅-C₈cycloalkyl, or 5- to 8-memberedheterocycloalkyl is independently optionally substituted by deuterium,halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, —OR^(e), —OC(O)R^(e),—OC(O)NR^(e)R^(f), —OC(═N)NR^(e)R^(f), —OS(O)R^(e), —OS(O)₂R^(e),—OS(O)NR^(e)R^(f), —OS(O)₂NR^(e)R^(f), —SR^(e), —S(O)R^(e), —S(O)₂R^(e),—S(O)NR^(e)R^(f), —S(O)₂NR^(e)R^(f), —NR^(e)R^(f), —NR^(e)C(O)R^(f),—NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(e)R^(f), —NR^(e)S(O)R^(f),—NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(e)R^(f), —NR^(e)S(O)₂NR^(e)R^(f),—C(O)R^(e), —C(O)OR^(e), —C(O)NR^(e)R^(f), —PR^(e)R^(f),—P(O)R^(e)R^(f), —P(O)₂R^(e)R^(f), —P(O)NR^(e)R^(f), —P(O)₂NR^(e)R^(f),—P(O)OR^(e), —P(O)₂OR^(e), —CN, or —NO₂; R⁶ is H, deuterium, or C₁-C₆alkyl, wherein each hydrogen atom in C₁-C₆ alkyl is independentlyoptionally substituted by deuterium, halogen, —OR^(e), —SR^(e), or—NR^(e)R^(f); each R⁷ is independently hydrogen or deuterium; eachR^(a), R^(b), R^(c), R^(d), R^(e), and R^(f) is independently selectedfrom the group consisting of H, deuterium, C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl,C₆-C₁₀ aryl, and 5- to 7-membered heteroaryl; and n is 0, 1, 2, or 3.19. The method of claim 18, wherein the subject is a human.